1、MX5301-1MG WSP-1 (H2S Probe) 硫化氢荧光探针
[1]Hu YM et.Reactive oxygen species-triggered off-on fluorescence donor for imaging hydrogen sulfide delivery in living cells.Chem. Sci., 2019,10, 7690-7694
WSP-1 (H2S probe) was obtained from Shanghai Maokang Biotechology Co., Ltd. To verify the intracellular ROS-triggered H2S release, a commercially available probe (WSP-1), whose fluorescence can be selectively turned on by H2S,17 was used for the detection of H2S in HeLa cells. As shown in Fig. S17 (images c–e),† the fluorescence of WSP-1 in the green channel rises with increasing the dose of NAB in the presence of H2O2, suggesting the generation of the gradually increased H2S.
[2]Zhao AS et al.Hydrogen sulphide-releasing aspirin enhances cell capabilities of anti-oxidative lesions and anti-inflammation.Med Gas Res. 2019 Jul-Sep;9(3):145-152. PMID: 31552879
HUVECs were cultured with each sample on coverslips for 24 hours, and then incubated with 250 μM H2S fluorescent probe WSP-1 (Maokangbio, Shanghai, China) for 30 minutes at 37o C away from light. After washing by PBS, cells were imaged at 476 nm by fluorescence microscope, and the fluorescent intensity was measured by Image J software.
[3]Xie Congkun et al.FeS@BSA Nanoclusters to Enable H2S‐Amplified ROS‐Based Therapy with MRI Guidance.Adv. Sci. 2020, 7, 1903512
WSP-1 (H2S probe) was purchased from Shanghai Maokang Bio Co., Ltd. Intracellular H2S detection: WSP-1 was used as intracellular H2S probe. Briefly, after treatment of cells with FeS@BSA nanoclusters (pH = 7.4 and pH = 6.0) for 6 h (PBS for control), WSP-1 (15 μM) was added, followed by incubation for 30 min. Subsequently, the cells were washed twice with PBS and observed under fluorescent microscope (Ex/Em:465/515 nm).
[4]Wang G et al.Zinc sulfide nanoparticle-decorated fibre mesh to enable localized H2S-amplified chemotherapy.Chem. Commun., 2020,56, 4304-4307
To investigate the intracellular production of H2S, a WSP-1 hydrosulfide probe was used following the standard protocol. In the presence of H2S, emission at 516 nm could be observed with the excitation at λ = 480 nm. WSP-1 hydrosulfide probe was obtained from Maokangbio(Shanghai, China)
「5」Ge Chunpo, et al.A novel NIR fluorescence probe with cysteine-activated structure for specific detection of cysteine and its application in vitro and in vivo.Talanta Volume 223, Part 2, 1 February 2021, 121758
NaHS andWSP-1were obtained from Aladain (Shanghai, China) andMaokang Biotechnology(Shanghai, China), respectively.
2、 MX4805-1MG Hydroxyphenyl fluorescein (HPF) 羟苯基荧光素
[1]Zhen WY et al.Reductive Surfactant-Assisted One-Step Fabrication of BiOI/BiOIO3 Heterojunction Biophotocatalyst for Enhanced Photodynamic Theranostics Overcoming Tumor Hypoxia.Nanoscale Horiz., 2019, 4, 720-726
Hydroxyphenyl fluorescein (HPF) was purchased from Shanghai Maokang Bio. Co. (Shanghai, China). HPF was used to detect the generation of •OH. Typically, 1 mL BB NCs aqueous solution (80 µg mL -1 ) containing with HPF (1 µM) were irradiated by 650 nm laser (0.5 W cm-2 ), the fluorescence of HPF was detected after 15 min. HPF aqueous solution with laser served as control.
[2]Liu Yang et al.Defect modified zinc oxide with augmenting sonodynamic reactive oxygen species generation.Biomaterials. Volume 251, August 2020, 120075
Hydroxyphenyl fluorescein (HPF) was purchased from Shanghai Maokang Bio. Co. (Shanghai, China).
[3]Xu X, Huang B, Zeng Z, Chen J, Huang Z, Guan Z, Chen M, Huang Y, Zhao C. Broaden sources and reduce expenditure: Tumor-specific transformable oxidative stress nanoamplifier enabling economized photodynamic therapy for reinforced oxidation therapy. Theranostics 2020; 10(23):10513-10530. doi:10.7150/thno.49731.
Hydroxyphenyl fluorescein (HPF) was purchased from Shanghai Maokang Bio. Co. (Shanghai, China). For detecting intracellular •OH generation, treated B16F10 cells were rinsed and stained with specific •OH probe HPF (10 μM in PBS) for 60 min, followed by rinsed with PBS and imaged using CLSM. To further confirm the Fe(II) consumption, after treatment with H2O2 (positive control), CA and HA@CA for 12 h, the cells were stained with a ferric ion probe, 3′,6′-bis (diethylamino)-2-(4-oxopent-2-en-2-ylamino) spiro (isoindoline-1,9′-xanthen)-3-one (10 μM) at 37 °C for 30 min. Finally, the treated cells were rinsed with PBS and observed by CLSM.
[4]Tang H, Li C, Zhang Y, et al.Targeted Manganese doped silica nano GSH-cleaner for treatment of Liver Cancer by destroying the intracellular redox homeostasis.Theranostics. 2020;10(21):9865-9887. Published 2020 Aug 2. doi:10.7150/thno.46771
Hydroxyphenyl fluorescein (HPF) was supplied by Shanghai Mao Kang biotechnology Co., Ltd. For evaluating the different types of ROS, DHE and HPF were selected as probes to assess the generation of intracellular •O2- and •OH, respectively. Briefly, HepG2 cells (5×105 cells per well) were seeded in a 6-well plate and incubated overnight. After that,Free SFB, MnMSN@SFB or FaPEG-MnMSN@SFB at different concentrations (10, 20, and 40 µg/mL of SFB equivalent) were added and treated for 6 h. Cells were then stained with DHE (20 µM) for another 45 min. Eventually, fluorescence microscope was employed to observe the fluorescence of cells at the wavelength of 535 nm. Moreover, to assess the generation of intracellular •OH, HepG2 cells (1×105 cells per well) were seeded into confocal dishes and incubated for 12 h. Then, Free SFB, MnMSN@SFB or FaPEG-MnMSN@SFB at different concentrations (10, 20, and 40 μg/mL of SFB equivalent) were added, meanwhile HPF (10 µM) was added to capture generative intracellular •OH, following by co-incubation for 6 h. Eventually, CLSM was used to observe the fluorescence of cells at 490 nm.
[5]Lili Zhang, Jiabao Huang, Shiqi Su, Xiaochun Wei, Lin Yang, Huanhuan Zhao, Jianqiang Yu, Jie Wang, Jiyun Hui, Shiya Hao, Shanshan Song, Yanyan Cao, Maoshuai Wang, Xiaowei Zhang, Yanyan Zhao, Zhiyong Wang, Weiqing Zeng, Hen-Ming Wu, Yuxiang Yuan, Xiansheng Zhang, Alice Y. Cheung, Qiaohong Duan,
FERONIA receptor kinase-regulated reactive oxygen species mediate self-incompatibility in Brassica rapa, Current Biology, 2021, ISSN 0960-9822,https://doi.org/10.1016/j.cub.2021.04.060.
Probe 20,70-Dichlorodihydrofluorescein diacetate (H2DCFDA), dihydroethidium (DHE), hydroxyphenyl fluorescein (HPF) are commonly used for ROS detection. HPF was used at 10 mM in 0.2 M PBS buffer (pH 6.1) for 2 hours. Comparable results were obtained in experiments tested with all three dyes, providing confidence for the observed changes in ROS levels.
[6]Mengsi Wu, Zhiyong Liu, Weian Zhang. An ultra-stable bio-inspired bacteriochlorin analogue for hypoxia-tolerant photodynamic therapy.Chemical Science2021,12(4) , 1295-1301.https://doi.org/10.1039/D0SC05525E
Singlet oxygen sensor green (SOSG), dihydroethidium (DHE), 3’-(4-hydroxyphenyl) fluorescein (HPF), Fetal bovine serum (FBS), Dulbecco’s modified Eagle’s medium (DMEM), Hoechst 33342 and 3-(4,5- dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Shanghai Maokang Biotechnology Co.,Ltd. Hydroxyl radical (•OH) was detected by fluorescence spectroscopy using hydroxyphenyl fluorescein (HPF), which reacts with •OH to produce high fluorescence emission at 530 nm. HPF was dissolved in DMF to gain 5 mM HPF solution. Then, 20 μL HPF was added into the solution of 2 mL FBC or FBC nanogels. The fluorescence changes were detected after illumination with 750 nm laser, and the excitation wavelength was 490 nm.
[8]Kinetics of the Photoelectron-Transfer Process Characterized by Real-Time Single-Molecule Fluorescence Imaging on Individual Photocatalyst Particles.
Jinghua An, Xiaoting Song, Wenbo Wan, Yanzheng Chen, Haibin Si, Huichuan Duan, Lu Li*, and Bo Taing*. Kinetics of the Photoelectron-Transfer Process Characterized by Real-Time Single-Molecule Fluorescence Imaging on Individual Photocatalyst Particles. ACS Catal. 2021, 11, 12, 6872–6882 Publication Date:May 27, 2021https://doi.org/10.1021/acscatal.1c00983
HPF and Amplex red from maokang biotechnology co. ltd.
3、 MX4804-1MG Aminophenyl fluorescein (APF) 氨基苯基荧光素
[1]Wu Q et al.Cascade enzymes within self-assembled hybrid nanogel mimicked neutrophil lysosomes for singlet oxygen elevated cancer therapy.Nat Commun. 2019 Jan 16;10(1):240.
The aminophenyl fluorescein (APF) as the HClO detector were purchased from Shanghai Maokangbio. The further EDT related therapeutic species including the ROS, 1O2 and the intermediate HClO have been detected in tumour tissues of different groups by using the corresponding fluorescent probes, DCFH-DA, SOSG and aminophenyl fluorescein (APF, as HClO detector).
[2]Juan J et al.Dynamic tracking of bulk nanobubbles from microbubbles shrinkage to collapse.Colloids and Surfaces A: Physicochemical and Engineering Aspects. Volume 589, 20 February 2020, 124430
3′- p – (aminophenyl) fluorescein (APF) was purchased from Shanghai Maokang Biotechnology Co., Ltd. (shanghai, China).
[3]Yahui Zhang, Weizhou Sha, Yang Liu, Wei Wang, Zhi Yuan. A facile composite nanoparticle promoted by photoelectron transfer and consumption for tumor combination therapy. Mater. Chem. Front., 2020,4, 3047-3056https://doi.org/10.1039/D0QM00447B
3′-(p-aminophenyl) fluorescein (APF) was purchased from maokang-bio (Shang hai).
[4]Li L, Shao C, Liu T, Chao Z, Chen H, Xiao F, He H, Wei Z, Zhu Y, Wang H, Zhang X, Wen Y, Yang B, He F, Tian L. An NIR-II-Emissive Photosensitizer for Hypoxia-Tolerant Photodynamic Theranostics. Adv Mater. 2020 Nov;32(45):e2003471. doi: 10.1002/adma.202003471. Epub 2020 Oct 7. PMID: 33029855.
2,7-Dichlorodi-hydrofluorescein diacetate (DCFH-DA), aminophenyl fluorescein (APF), and singlet oxygen sensor green (SOSG) were obtained from Shanghai Maokang Biotechnology Co., Ltd. (Shanghai, China).
4、 MS4040-1G Fluorescent Brightener 28 (Calcofluor White M2R) 荧光增白剂28
[1]Yu C et al.Nanoprobe-based force spectroscopy as a versatile platform for probing the mechanical adhesion of bacteria. Nanoscale.2019 Apr 23;11(16):7648-7655. PMID: 30720812
Fluorescent Brightener 28 wasobtained from Maokang Biotechnology (Shanghai, China). For fluorescence staining of extracellularpolymeric substances (EPSs), after the biofilm formation, thePDMS surfaces were incubated with 6 µM SYTO 9 at roomtemperature in the dark for 12 min, followed by a second stain-ing of 0.15 mM fluorescent Brightener 28 for 3 min.
[2]Shengnan Tao et al. Ultra-stable Pickering emulsion stabilized by a natural particle bilayer. Chem. Commun., 2020,56, 14011-14014https://doi.org/10.1039/D0CC05690A
Calcofluor White was bought from Maokang Biotechnology (China) Co., Ltd. Calcofluor White (SNCs) and Nile Blue A (ZNPs) were used to stain Pickering emulsions. Lasers of 405 nm and 633 nm were used to excite the dyes of Calcofluor White and Nile Blue A, respectively.
[3]Qian Li et al. 2‐Hydroxy‐4‐methoxybenzaldehyde inhibits the growth ofAspergillus flavusvia damaging cell wall, cell membrane, manipulating respiration thus creating a promising antifungal effect on corn kernels.https://doi.org/10.1111/ijfs.14617
Calcofluor-White Stain (CAS: 4404-43-7) was purchased from Maokang Biotechnology Co. (Shang-hai, China).
[4]Ya Fei Geng, et al. An innovative role for luteolin as a natural quorum sensing inhibitor in Pseudomonas aeruginosa,Life Sciences,Volume 274,2021,119325,ISSN 0024-3205,https://doi.org/10.1016/j.lfs.2021.119325.
Afterwards, 50 μg/mL of Calcofluor white M2R (Maokang, Shanghai, China), a blue fluorescent marker, was used to visualize the exopolysaccharides in the biofilm matrix by staining for 2 h at room temperature in the dark.
[5] Li, Q., Zhu, X., Xie, Y.et al.o-Vanillin, a promising antifungal agent, inhibitsAspergillus flavusby disrupting the integrity of cell walls and cell membranes.Appl Microbiol Biotechnol105,5147–5158 (2021). https://doi.org/10.1007/s00253-021-11371-2
Calcofluor white (CAS: 4404-43-7) was purchased from Maokang Biotechnology Co. (Shanghai, China)
[6] JiashunShi et al. Constructing zwitterionic nanofiber film for anti-adhesion of marine corrosive microorganisms. Journal of Materials Science & Technology. Volume 70, 20 April 2021, Pages 145-155
Calcofluor stain was purchased from Maokang Biotechnology Co., Ltd (Shanghai, China).
[7] QianLi et al. Antifungal efficacy of paeonol on Aspergillus flavus and its mode of action on cell walls and cell membranes. LWT Volume 149, September 2021, 111985
Calcofluor white (CW) (CAS: 4404-43-7) were purchased from the Maokang Biotechnology Co., Ltd
Fig. 1. Effect of paeonol on cell walls’ integrity ofA. flavus. (A) SEM observation of the mycelial morphology. Quantification of the content of (B) β-1,3-glucan and (C) chitin in mycelia cell walls. a-c significant difference (P< 0.05) according to Duncan’s multiple range test. (D) Distribution of septa in mycelia treated with paeonol after staining with calcofluor white under a fluorescence microscope. Scale bar: 10 μm.
https://ifst.onlinelibrary.wiley.com/doi/abs/10.1111/ijfs.14617
5、 MX7356-500MG Lywallzyme 真菌溶壁酶
Jin W et al.Importance of a Laccase Gene (Lcc1) in the Development of Ganoderma tsugae.Int J Mol Sci. 2018 Feb 6;19(2). PMID: 29415422
Finally, with gentle agitation, the mycelia (approximately 0.2 g) were incubated for 4–5 h at 31 °C in 2 mL of 20 mg/mL lywallzyme (Shanghai Maokang Biological Technology Co, Ltd., China, Catalog No. MX7365-500MG) containing 0.6 M mannitol for preparing protoplasts. After incubation, these protoplasts were washed free of enzyme mannitol solution and transferred to regeneration medium (RM).
6、MZ2103-200UL Brefeldin A (BFA, 5mg/ml)
Zeng C et al.Th17 cells were recruited and accumulated in the cerebrospinal fluid and correlated with the poor prognosis of anti-NMDAR encephalitis. Acta Biochim Biophys Sin (Shanghai).2018 Dec 1;50(12):1266-1273.
Cells were pretreated with 25 ng/ml PMA (Sigma, St Louis, USA) for 5 h at 37°C. At the same time, 10 μg/ml brefeldin (Maokang Biotechnology, Shanghai, China) was added into the cells for the last 4 h of the incubation.
7、 MX4509-100UG SBFI AM ( Na+ Indicator) 钠离子指示探针
MX4510-100UG PBFI AM ( K+ Indicator) 钾离子指示探针
[1]Tong W et al.Phthalocyanine functionalized poly(glycidyl methacrylate) nano-assemblies for photodynamic inactivation of bacteria.Biomater. Sci., 2019,7, 1905-1918
Sodium-binding benzofuran isophthalate acetoxymethyl ester (SBFI-AM) and Potassium-binding Benzofuran Isophthalate Acetoxymethyl ester (PBFI-AM) were purchased from MKbio. China.
[2]Li R et al.Biofilm inhibition and mode of action of epigallocatechin gallate against Vibrio mimicus.Food Control, Volume 113, July 2020, 107148
Then PBFI probe (ShangHai MaoKang Bio technology Co., LTD., China) was added and incubated at 37 °C for 90 min. The cells were washed, collected and resuspended with PBS buffer. Aliquots (100 μL) of bacterial suspension were transferred to a Corning 96 well black plate, and 100 μL of various concentrations of EGCG were dispensed in the microtiter plate wells.
[3]Liu Y, Zhen W, Wang Y, Song S, Zhang H. Na2S2O8Nanoparticles Trigger Antitumor Immunotherapy through Reactive Oxygen Species Storm and Surge of Tumor Osmolarity. J Am Chem Soc. 2020 Dec 30;142(52):21751-21757. doi: 10.1021/jacs.0c09482. Epub 2020 Dec 18. PMID: 33337859.
4T1 cells were inoculated into glass bottom culture dishes for 24 h. Then, adding PNSO NPs medium solution (80 μg/mL) to continue co-culture for 4 h. The treated 4T1 cells were further incubated with 10
µM Na+ indicator SBFI AM (sodiumbinding benzofuran isophthalate acetoxymethyl ester, MKBio, 90%) in 0.04% Pluronic F-127 (MKBio) and the fluorescence signal was measured by CLSM.
[4] Liang Z, Yang Y, Yu G, et al. Engineering aluminum hydroxyphosphate nanoparticles with well-controlled surface property to enhance humoral immune responses as vaccine adjuvants. Biomaterials. 2021 Jun;275:120960. DOI: 10.1016/j.biomaterials.2021.120960.
BMDMs were treated with AAHPs (250 μg/mL) in the presence of LPS at 500 ng/mL for 5 h. Then PBFI AM (Mkbio, Shanghai, China) was added to the cells at the concentration of 10 μM, and cells were incubated at 37 °C for 1h. Triton X-100 (0.2%) treated cells were used as controls. The fluorescence of PBFI AM was measured at the Ex/Em of 340/615 nm. The data were expressed as relative fluorescence intensity (RFI) defined as the fluorescence intensity of AAHPs-treated BMDMs normalized to the intensity of control cells.
8、MP6101-25G BSA, Standard Grade 牛血清白蛋白（标准级别）
Ma MQ et al.Nanocomposite membranes embedded with functionalized MoS2 nanosheets for enhanced interfacialcompatibility and nanofiltration performance.Journal of Membrane Science Volume 591, 1 December 2019, 117316
Bovine serum albumin (BSA, Standard Grade) was acquired from Shanghai Maokang Biotechnology Co., Ltd (China). Polyacrylonitrile ultrafiltration membranes (PAN, MWCO ranging from 10 to 300 kDa) were purchased from Shanghai MegaVision Membrane Engineering & Technology Co., Ltd (China).
9、 MX3008-5ML Cell Counting Kit (CCK-8)
Liu JY et al.3D printing of biomimetic multi-layered GelMA/nHA scaffold for osteochondral defect repair.Materials & Design Volume 171, 5 June 2019, 107708
Cell counting kit-8 reagent was purchased from Shanghai Maokang Biotechnology Co., Ltd. (China). Cell proliferation on the scaffold was measured with CCK-8 after 1, 3, 5 and 7 days of culture, and cell culture plates were used as blank group. The BMSCs-seeded hydrogels were incubated in DMEM containing 10% CCK-8 protected from light at an incubator of 37 °C and 5% CO2 for 4 h, and measured at 450 nm using a microplate reader.
10、 MX3012-500T Calcein AM/PI Double Stain Kit 活细胞/死细胞双染试剂盒
Zhang XX et al.Characterization and property of dual-functional Zn-incorporated TiO2 micro-arc oxidation coatings: The influence of current density.Journal of Alloys and Compounds Volume 810, 25 November 2019, 151893
After the formazan was dissolved by dimethyl sulfoxide (DMSO), the optical density (OD) was then determined using enzyme-labeled instrument (iMark, BIO-RAD, US) at the wavelength of 570 nm. To reveal the viability of adherent MC3T3-E1 cells, a Live/Dead Double Staining Kit (MKBio, China).
Double Staining Kit (MKBio, China) was employed. After 1 day culture, … stained with Calcein acetoxymethyl ester (Calcein AM) and propidium.
11、 MX4801-1KIT Reactive Oxygen Species (ROS) Assay Kit 活性氧检测试剂盒
[1]Wang YW et al.Screening of Duck Tembusu Virus NS3 Interacting Host Proteins and Identification of Its Specific Interplay Domains.Viruses 2019, 11(8), 740;
HEK293 cells were first transfected with pCAGGS and pCAGGS-PRDX1. When PRDX1 was highly expressed for about 24 h, cells were infected with DTMUV and treated for 6 h, whereas the Rosup positive control group was set in parallel. Finally, the probe of DCFH-DA was diluted at 1:1000 and treated at 30 °C for 20 min referring to the ROS Assay Kit (MKBio, Shanghai, China). Cells were then observed under a fluorescent inverted microscope immediately.
[2]Zhi-Hao Hu, Guo-Jun Wang, Rui-Xin Li, Tian-Yu Zhu, Zhuo-Yin Wang, Heng-Xuan Ding, Xiu-Mei Hu, Upregulation of miR-133a-3p enhances Bufothionine-induced gastric cancer cell death by modulating IGF1R/PI3K/Akt signal pathway mediated ER stress, Life Sciences, 10.1016/j.lfs.2020.118180, (118180), (2020).
The DCFH-DA ROS assay kit (MKBio, Shanghai, China) was used to measure the ROS levels of the GC cells. In brief, GC cells were prepared and incubated with 5 μM DCFH-DA staining solution in the dark for 30 min at 37 °C.
[3]Liu, Y.; Xiao, S.; Sun, H.; Pei, L.; Liu, Y.; Peng, L.; Gao, X.; Liu, Y.; Wang, J. AtPPRT1, an E3 Ubiquitin Ligase, Enhances the Thermotolerance in Arabidopsis. Plants 2020, 9, 1074.
The ROS production in the seedlings was detected by using fluorescent dye 2′,7′-dichlorofluorescein diacetate (H2DCFDA) (Maokang, Shanghai, China), as described previously [32]. The 7-day-old different-genotype seedlings grown on MS solid medium were exposed to 45 °C for 3 h, and the seedlings were collected and stained at 22 °C for 15 min in 50-μM H2DCFDA (dissolved in MES-KCl buffer, MES 10 mM, KCl 50 mM, adjust to pH 5.5 with HCl). The seedlings washed with distilled water were placed under a Leica confocal microscope. The fluorescence in cotyledons and roots of seedlings were detected by using an excitation wavelength of 488 nm
[4]Shen LD, Qi WH, Bai JJ, Zuo CY, Bai DL, Gao WD, Zong XL, Hao TT, Ma Y, Cao GC. Resibufogenin inhibited colorectal cancer cell growth and tumorigenesis through triggering ferroptosis and ROS production mediated by GPX4 inactivation. Anat Rec (Hoboken). 2021 Feb;304(2):313-322. doi: 10.1002/ar.24378. Epub 2020 Feb 7. PMID: 31961485.
The intracellular ROS levels in CRC cells were measured by using the
2,7-dichlorodihydrofluorescein diacetate (DCFH- DA) ROS assay kit purchased from MKBio
(Shanghai, China) according to the manufacturer’s instruction
[5]Yan, W., Zhang, Y., Hu, L.et al.Febuxostat Inhibits MPP+-Induced Inflammatory Response Through Inhibiting the JNK/NF-κB Pathway in Astrocytes.Neurotox Res39,566–574 (2021).https://doi.org/10.1007/s12640-020-00316-8
The astrocytes were planted on 24-well cell culture plates at a density of 1 × 10 5 cells/well for 24 h. After necessary stimulation, cells were loaded with 5 μM DCFH-DA (MX4802, MKBio, China) and incubated for 30 min in darkness.
12、MP1509-25KU Benzonase Nuclease 全能核酸酶（无内毒素）
Raihan O et al.SFRS11 Loss Leads to Aging-Associated Cognitive Decline by Modulating LRP8 and ApoE.Cell Rep. 2019 Jul 2;28(1):78-90.e6.
AAV particles were released from cells with three freeze/thaw cycles, followed by incubation with 50 U/ml benzonase nuclease (MKbio) and 10 U/ml RNase I at 37°C for 30 min, and 0.5% sodium deoxycholate (Sigma) treatment for another 30 min.
13、 MZ2151-1MG Ionomycin, Free Base 离子霉素（游离酸）
Xu Z et al.Immunomodulatory effects of Rhipicephalus haemaphysaloides serpin RHS2 on host immune responses.Parasit Vectors. 2019 Jul 11;12(1):341.
Ionomycin (Free Base) was purchased from MKbio (Shanghai, China). Recombinant mouse GM-CSF (granulocyte-macrophage colony stimulating factor), IL-4 (interleukin 4) and IL-2 (interleukin 2) were obtained from Peprotech (Rocky Hill, NJ, USA).
14、MS4022-100MG Nile Red 尼罗红
Shi Y et al.Increasing the removal of protein-bound uremic toxins by liposome-supported hemodialysis. Artif Organs.2019 May;43(5):490-503.
15、MP6308-100UG FITC-UEA-I (FITC labeled Ulex Europaeus Agglutinin I)
MP6013-500UG Human DiI-Ac-LDL
[1]Quan H et al.LncRNA-AK131850 Sponges MiR-93-5p in Newborn and Mature Osteoclasts to Enhance the Secretion of Vascular Endothelial Growth Factor a Promoting Vasculogenesis of Endothelial Progenitor Cells.Cell Physiol Biochem. 2018;46(1):401-417.
After washing, cells were plated on culture dishes at the density of 5×10 6 and cultured with Microvascular Endothelial Cell Growth Medium-2 (EGM-2 MV) BulletKit (Lonza, Walkersville, MD, USA) in a humidified atmosphere of 5% CO2 at 37°C. Two days later, non-adherent cells were removed by washing with PBS twice and the culture medium was replaced every 3 d. After culturing for 1 d, 7 d, 14 d and 28 d, EPCs were identified by optical microscope observation, immunofluorescence for both Dil-Acetylated Low Density Lipoprotein (Dil-Ac- LDL; MaoKang Biotechnology, Shanghai, China) and FITC labeled Ulex Europaeus Agglutinin 1 (FITC-UEA-1; MaoKang Biotechnology, Shanghai, China), and flow cytometry (FACSCalibur Flow Cytometry, BD, Triangle, NC, USA) for PE-Cy7 conjugated anti-mouse CD34 monoclonal antibody.
[2]Li Li et al.Dual-Peptide-Functionalized Nanofibrous Scaffolds Recruit Host Endothelial Progenitor Cells for Vasculogenesis to Repair Calvarial Defects.ACS Applied Materials & Interfaces 2020 12 (3), 3474-3493 DOI: 10.1021/acsami.9b21434
acetylated low-density lipoprotein (DiI-ac-LDL; Maokang, China) in EGM-2 medium for 4 h at 37 °C. Cells were fixed in 4% paraformaldehyde
[3]Wang, W., Zhang, Y., Hui, H.et al.The effect of endothelial progenitor cell transplantation on neointimal hyperplasia and
eendothelialization after balloon catheter injury in rat carotid arteries.Stem Cell Res Ther12,99 (2021).https://doi.org/10.1186/s13287-021-02135-w
During the 8 days of culture, attached EPCs were washed with PBS and subsequently stained with 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine-labelled acetylated low-density lipoprotein (Dil-ac-LDL, Maokang Biotechnogy Co., Ltd., Shanghai, China, MP6013) at a concentration of 10 mg/L. After incubation at 37 °C and 5% CO2for 4 h, cells were fixed with 2% paraformaldehyde (PFA) in PBS for 10 min. Fluorescein isothiocyanate-labelledUlex europaeusagglutinin (FITC-UEA-1, Maokang Biotechnogy Co., Ltd., Shanghai, China, MP6308) at a concentration of 10 mg/L was added. The cells were examined under a confocal fluorescent microscope (TCS SP5 II, Leica Microsystems, Germany).
16、MP7501-100T Alkaline Phosphatase Assay Kit (Colorimetric)
Wang CH et al.Colorimetric logic gate for alkaline phosphatase based on copper (II)-based metal-organic frameworks with peroxidase-like activity.Analytica Chimica Acta Volume 1004, 3 April 2018, Pages 74-81
Alkaline Phosphatase Colorimetric Assay Kit using p-nitrophenyl phosphate (pNPP) as aphosphatase substrate was purchased from MKbio Co. Ltd. (Shanghai, China).
17、MX2201-1ML Polybrene (10mg/ml)
Dai XJ et al.AHIF promotes glioblastoma progression and radioresistance via exosomes.Int J Oncol. 2019 Jan;54(1):261-270.
The cells were then infected with the same virus titer on the following day with. 8 µg/ml Polybrene (Maokang Co., Shanghai, China). At ~72 h.
18、MX4021-100UL PKH26 Cell Linker Kit for General Cell Membrane Labeling PKH26 细胞连接试剂盒（用于常规细胞膜标记）
Yu Mengyu et al.Targeted exosome‐encapsulated erastin induced ferroptosis in triple negative breast cancer cells.Cancer Science. 2019;110:3173–3182.
Targeted exosome‐encapsulated erastin induced ferroptosis in triple negative breast cancer cells.
To quantify the amount of erastin@FA‐exo and erastin@exo taken up by MDA‐MB‐231 cells, lipophilic fluorescent dye PKH26 (MaoKang Biotechnology) was used to stain the exosomes. To detect the effect of FA receptor binding on cell uptake, culture medium containing 1.1 mg/mL of free FA was added to MDA‐MB‐231 cells to competitively inhibit FA receptors. After incubation for 6 hours, the cells were washed with PBS 3 times.Then erastin@FA‐exo was added and the cells’ uptake of the drug was observed.
19. MX4008-100MG DiOC2(3) 绿色膜电位荧光探针
[1]Chen Z et al.Hypoionic Shock Facilitates Aminoglycoside Killing of Both Nutrient Shift- and Starvation-Induced Bacterial Persister Cells by Rapidly Enhancing Aminoglycoside Uptake. Front Microbiol. 2019 Sep 6;10:2028. PMID: 31551965
A flow cytometry-based assay was applied to measure the PMF by using the fluorescence probe 3,3′-Diethyloxacarbocyanine Iodide [DiOC2(3); purchased from MaoKang Biotechnology, Inc., Shanghai, China] according to the manufacturer’s instruction. Briefly, E. coli persisters, with or without CCCP pretreatment as described above, were diluted into PBS to a cell density of 106 cells/mL and incubated with DiOC2(3) (at a final concentration of 30 μM) at room temperature for 30 min. Cells were subjected to flow cytometric analysis on FACSymphonyTMA5 (BD Biosciences) with an excitation at 488 nm and emission at both red and green channels.
[2]Zhao Y et al.Rapid Freezing Enables Aminoglycosides To Eradicate Bacterial Persisters via Enhancing Mechanosensitive Channel MscL-Mediated Antibiotic Uptake.mBio. 2020 Feb 11;11(1). pii: e03239-19. doi: 10.1128/mBio.03239-19. PMID: 32047133
A flow cytometry-based assay was applied to measure the proton motive force by using the fluorescence probe 3,3′-diethyloxacarbocyanine iodide [DiOC2(3)] (purchased from MaoKang Biotechnology, Inc., Shanghai, China) according to the manufacturer’s instructions. Briefly, E. coli exponential-phase cells, with or without CCCP pretreatment for 1 h, were diluted into PBS to a cell density of 106 cells/ml and incubated with DiOC2(3) (at a final concentration of 30 μM) at room temperature for 15 min. Cells were subjected to flow cytometric analysis on a FACSymphony A5 system (BD Biosciences) with excitation at 488 nm and emission in both the red (630-nm) channel and green (515-nm) channel. Cells treated by rapid freezing for 10 s with liquid nitrogen were also analyzed.
20、MS4328-250G PVP-40 (Polyvinylpyrrolidone, average MW 40,000) 聚乙烯吡咯烷酮
Ying-Ya Li, Yan-Ping Jia, Li-Yan Duan, and Kun-Ming Li.Participation of the inositol 1,4,5-trisphosphate-gated calcium channel in the zona pellucida- and progesterone-induced acrosome reaction and calcium influx in human spermatozoa.Asian J Androl. 2020 Mar-Apr; 22(2): 192–199.
Hoechst 33258 and polyvinylpyrrolidone-40 (PVP-40) were obtained from Shanghai Maokang Biotechnology Co., Ltd. (Shanghai, China). The spermatozoa were washed and incubated with Hoechst 33258 (4 μg ml−1) for 10 min at a proportion of 9:1 to achieve a final concentration of 0.4 μg ml−1 and then centrifuged with 2% (w/v) PVP-40 in PBS. The supernatant was then removed and resuspended in PBS, smeared on clean slides, and fixed for 30 min in 95% (v/v) ethanol after air drying.
21、MP7505-30ML Alkaline Phosphatase Stain Solution (Coupling azo-dye Method) 碱性磷酸酶染色液（偶氮偶联法）
[1]Fu, L., Jin, P., Hu, Y., Lu, H., & Su, L. (2020).KR‑12‑a6 promotes the osteogenic differentiation of human bone marrow mesenchymal stem cells via BMP/SMAD signaling.Molecular Medicine Reports, 21, 61-68.
For ALP staining, hBMSCs in each group were washed twice with PBS at the end of the 7 day KR 12 a6 treatment period, fixed with 4% paraformaldehyde for 30 s, and then stained using the ALP staining kit (Maokang Biotechnology, Shanghai) according to the manufacturer’s instructions.images were acquired by an inverted phase contrast microscope (CKX41, Olympus) under x100 magnification. At least five fields per sample were randomly selected and observed.
[2]Peiying Lv, Pengfei Gao, Yanyan Yang, Zihui Wang, Lu Sun, Feifei Mo, Guangjie Tian, Mingjie Kuang, Yonglan Wang.Osteocyte-derived exosomes induced by mechanical strain promote human periodontal ligament stem cell proliferation and osteogenic differentiation in an inflammatory environment via the miR-181b-5p/PTEN/AKT signaling pathway.Stem Cell Research & Therapy；
Osteogenic Differentiation Assay: HPDLSCs in different groups (2 × 105 cells/well) were cultured in 6-well plates containing osteogenic differentiation medium (Cyagen, Suzhou, China). The medium was refreshed every 3 days until the 21st day. Cells were fixed with 4% paraformaldehyde. Mineralized nodules were observed by staining with a 1% Alizarin red solution (Cyagen, Suzhou, China). The activity of alkaline phosphatase (ALP) was analyzed using an Alkaline Phosphatase Staining Kit (MKbio, Shanghai, China). Staining images were acquired using an inverted microscope, and ImageJ was used to analyze the images.
22、Puromycin Dihydrochloride 嘌呤霉素盐酸盐MS0011-25MG
[1]Li, X., Tang, M., Zhu, Q. et al.The exosomal integrin α5β1/AEP complex derived from epithelial ovarian cancer cells promotes peritoneal metastasis through regulating mesothelial cell proliferation and migration.Cell Oncol. (2020).
SKOV3 cells were transfected with a luciferase reporter vector containing a puromycin resistance gene (SKOV3-luc) and grown in complete medium supplemented with 3 μg/ml puromycin (Shanghai MaoKang Biotechnology, MS0011-25MG). The cells were maintained in culture for no more than 10 passages and they were authenticated using short tandem repeat (STR) analysis every six months.
[2]Zhou J et al.Exosomes Released from Tumor-Associated Macrophages Transfer miRNAs That Induce a Treg/Th17 Cell Imbalance in Epithelial Ovarian Cancer.Cancer Immunol Res December 1 2018 (6) (12) 1578-1592; DOI: 10.1158/2326-6066.CIR-17-0479
The original cell culture medium was then removed, and the lentivirus was added to the cells and incubated overnight. After 24 hours of infection, the culture medium containing the lentivirus was removed and replaced with fresh culture medium. After 72 hours, the transfected cells were grown in complete high-glucose DMEM with puromycin (3 μg/mL; Shanghai MaoKang Biotechnology, MS0011-25MG) for purity screening.
[3]Chen X et al.Exosomes derived from hypoxic epithelial ovarian cancer cells deliver microRNAs to macrophages and elicit a tumor-promoted phenotype.Cancer Lett. 2018 Oct 28;435:80-91. doi: 10.1016/j.canlet.2018.08.001. Epub 2018 Aug 8.
The SKOV3 cell line was transfected into a luciferase reporter along with puromycin resistance gene (SKOV3-Luc-pur), which was grown in RPMI-1640 with 10% FBS, Gibco, 1% antibiotics (Gibco), and 3 μg/ml puromycin (Shanghai MaoKang Biotechnology, MS0011-25 MG) for purity screening.
23、MX4812-5MG Dihydroethidium (DHE) 二氢乙锭
Zhao AS et al.Hydrogen sulphide-releasing aspirin enhances cell capabilities of anti-oxidative lesions and anti-inflammation.Med Gas Res. 2019 Jul-Sep;9(3):145-152. PMID: 31552879
HUVECs and macrophages were respectively seeded on coverslips and cultured with samples for 24 hours and then treated with 400 μM H2O2 for 12 hours. After washing by PBS, cells were incubated with 10 μM ROS fluorescent probe-dihydroethidium (Maokangbio, Shanghai, China) at 37o C for 30 minutes away from light. Solution was removed for detection. Cells were observed and photographed at 488 nm using fluorescence microscope, with the fluorescent intensity measured by ImageJ software.
24、MM1506-500ML4% paraformaldehyde 4%多聚甲醛
Cell invasion was detected using transwell chamber (Corning, NY, USA). Briefly, cells were seeded in the upper chamber pre-coated with matrigel at a density of 1 × 105/chamber. The medium containing 10% FBS was added in the lower chamber. After 24 h of culturing, cells on the upper chamber were removed. Cells in the lower chamber were fixed with 4% paraformaldehyde (Shanghai Maokang Biotechnology Co., Ltd, Shanghai, China) for 15 min, and stained with 0.1% crystal violet for 30 min. Positive stained cells were counted under a microscope (Olympus) at five randomly selected fields, and the invasion rate was calculated.
25、 MX4403-300T FITC Phalloidin FITC标记鬼笔环肽
[1]Qian Xu et al.Novel injectable and self-setting composite materials for bone defect repair.Download PDF SCIENCE CHINA Materials, Volume 63, Issue 5: 876-887(2020)
MC3T3-E1 cells were first inoculated into six-well plates at the initial density of 4×104 cells/well (4 mL per well) for 24 h. The cell medium was replaced by 4 mL of the extraction medium when the cell confluence reached approximately 80%. Cells were gently washed by PBS and fixed with 4% polyformaldehyde for 20 min, followed by PBS washing for three times. A fluorescein phalloidin (
) solution (10 µg mL−1) (Mao Kang Biotechnology Co., Ltd., Shanghai, China) was added to the wells and stained for 50 min. The samples were subsequently washed with PBS three times and stained with a 4ʹ,6-diamidino-2-phenylindole (DAPI) solution (100 ng mL−1) to counterstain the cell nuclei for 15 min. These samples were finally washed with PBS before observation under an Eclipse Ti inverted fluorescence microscope (Nikon Instruments Inc., Tokyo, Japan).
[2]Han Y, Lian M, Sun B, Jia B, Wu Q, Qiao Z, Dai K.Preparation of high precision multilayer scaffolds based on Melt Electro-Writing to repair cartilage injury.Theranostics 2020; 10(22):10214-10230. doi:10.7150/thno.47909.
After 21 days of culturing, the cell-bearing scaffolds (n = 4 per group) were fixed in 4% paraformaldehyde for 2 h. Triton-X (0.1%) was used to soak the scaffolds for 5 min, which were then washed thrice with PBS for 5 min each. Blocking was performed with BSA at room temperature for 1 h. After blotting with water-absorbing paper, primary antibodies (anti-PRG4, anti-CILP, anti-COLII, anti-COLI, and anti-SOX-9) (1:100, Abcam, UK) were added to each sample and incubated at 4 °C overnight. Following washes with PBS, a corresponding secondary antibody (1: 500, Abcam, UK) was added, incubated at room temperature for 1 h, and washed with PBS. The scaffolds were then labeled with FITC phalloidin (1:200, MaoKang, China) and incubated at room temperature for 2 h, followed by incubation with DAPI staining solution (1:1000, MaoKang, China) for 30 min at room temperature. After the final wash with PBS, samples were observed and imaged under a confocal microscope (LEICA, Germany). Analysis and quantification of protein expression was performed using ImageJ.
26、MX4511-1MG 2-NBDG 葡萄糖摄取荧光探针
Zhang Yh et al.A New Possible Mechanism by Which Punicalagin Protects against Liver Injury Induced by Type 2 Diabetes Mellitus: Upregulation of Autophagy via the Akt/FoxO3a Signaling Pathway.J. Agric. Food Chem. 2019, 67, 50, 13948–13959
2-NBDG was purchased from Shanghai maokang biotechnology co.,ltd
27、MX5302-1MG WSP-5 (H2S Probe) 硫化氢荧光探针
He T et al.Tumor pH-responsive metastable-phase manganese sulfide nanotheranostics for traceable hydrogen sulfide gas therapy primed chemodynamic therapy.Theranostics. 2020; 10(6): 2453–2462.
H2S Fluorescent Probe WSP-5 was purchased from Shanghai Maokang Biotechnology
Co., Ltd. (Shanghai, China). H2S gas released in 4T1 cells was observed by adding a Washington State Probe-5 (WSP-5) H2S fluorescent probe (Figure Figure33H). It is based on 2-pyridyl disulfide fluorescent, which release the fluorophores and turn on the fluorescence by tandem nucleophilic subsitution-cyclization reaction. WSP-5 probe can selectively and rapidly react with H2S in cells to generate a green fluorescence signal
28、MX3006 Alamar Blue Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
Wang Lixuan et al.Fabrication of Injectable, Porous Hyaluronic Acid Hydrogel Based on an In-Situ Bubble-Forming Hydrogel Entrapment Process.Polymers 2020, 12(5), 1138;https://doi.org/10.3390/polym12051138
Alamar Blue was bought from Maokang Biotechnology Co., Ltd., (Shanghai, China). To detect cytotoxicity and cell proliferation, the rBMSC suspension was added to the wells from the top of the scaffold, and culturing took place in a cell incubator at 37 °C for 1, 4, or 7 days. After the end of the culturing, we added Alamar blue reagent to the well plate and continued incubating in the cell incubator for 2–8 h until the color of the culture medium changed from blue to pink. Finally, the fluorescence intensity which is directly proportional to the number of viable cells was measured by a fluorescence photometer (SAFIRE, Tecan Co., Mannedorf, Switzerland).
29、MS1601 Streptozotocin (STZ) 链脲佐菌素
YAN, Wensheng; JIANG, Lingjun and XU, Jifen.Cyclocarya paliurus (Batal.) Iljinskaja polysaccharides alleviate type 2 diabetes mellitus in rats by resisting inflammatory response and oxidative stress.Food Sci. Technol [online]. 2020, vol.40, suppl.1 [cited 2020-07-24], pp.158-162.
Streptozotocin was provided by Shanghai Maokang Biotechnology Co., Ltd. (Shanghai, China). Sixty rats were single-cage raised in the condition avoiding strong light and noise (temperature 22 ± 2 °C; humidity 40-70%; 12/12-h day-night cycle; free to feed and water). After one week of adaptive feeding, 10 rats were randomly selected as the control group, which were fed with standard diet. The remaining 50 rats were fed with high-fat and high-sugar diet (20% sucrose, 18% lard, 3% yolk and 59% basic diet) for 4 weeks. The rats were fasted for 12 h, followed by single intraperitoneal injection of streptozotocin solution with dose of 30 mg/kg. After 72 h, the blood sample was taken from tail vein, and the fasting blood glucose (FBG) was detected. The FBG > 16.7 mmol/L presented the successful establishment of T2DM model.
30、 MX5202-1MG ROSGreenTM H2O2 Probe 过氧化氢荧光探针
[1]Ma W, Chen X, Fu L, et al.Ultra-fficient Antibacterial System Based on Photodynamic Therapy and CO Gas Therapy for Synergistic Antibacterial and Ablation Biofilms.ACS Appl Mater Interfaces. 2020;12(20):22479-22491. doi:10.1021/acsami.0c01967
Variation of ROS and H2O2 level in E. coli after CO release was respectively measured by DCFH-DA (10 μΜ) and ROSGreenTM H2O2 Probe (5 μM) using the parallel method. The λEx and λEm for DCFH-DA were 505 and 535 nm, respectively, while that for ROSGreenTM H2O2 Probe were 490 and 525 nm, respectively. DCFH-DA and ROSGreenTM H2O2 Probe were purchased from Maokang Co., Ltd. (Shanghai,China).
[2]Luo X, Wang R, Lv C, Chen G, You J, Yu F.Detection of Selenocysteine with a Ratiometric near-Infrared Fluorescent Probe in Cells and in Mice Thyroid Diseases Model.Anal Chem. 2020;92(1):1589-1597. doi:10.1021/acs.analchem.9b04860
Besides, both of our probes Mito-Cy-Sec and commercial ROSGreen H2O2 are employed to examine the interrelationship between H2O2 and Sec in cells and in mice models. The results demonstrate that the relevant-levels between H2O2 and Sec are exactly negative correlation.
31、MX4005 DiR Iodide (DiIC18(7))
[1]Cai J, Qian K, Zuo X, et al.PLGA nanoparticle-based docetaxel/LY294002 drug delivery system enhances antitumor activities against gastric cancer.Journal of Biomaterials Applications. 2019;33(10):1394-1406. doi:10.1177/0885328219837683
The in vivo tumor-targeting capacity of the NPs was examined using a lipophilic carbocyanine dye DiR (MaokangBio, Shanghai, China).
32、MS0905-0100MG FITC-Dextran 70 (FD70) FITC标记葡聚糖（70 kDa）
Tang, X., Yan, K., Wang, Y.et al.Activation of PPAR-β/δ Attenuates Brain Injury by Suppressing Inflammation and Apoptosis in a Collagenase-Induced Intracerebral Hemorrhage Mouse Model.Neurochem Res45,837–850 (2020). https://doi.org/10.1007/s11064-020-02956-w
BBB permeability was also evaluated using 70 kDa FITC-dextran. Mice (n = 5/group) were injected with 70 kDa FITC-dextran (MS0905; MKbio, Shanghai, China) in PBS at 50 mg/kg into the tail vein. Animals were perfused with pre-chilled PBS 15 min after the dextran injection. Following perfusion, the brain tissue was removed, homogenized with 20% trichloroacetic acid, and centrifuged at 12,000 rpm for 10 min. The supernatant was isolated, and relative fluorescence was measured at excitation and emission wavelengths of 493 and 520 nm. The results were calculated using FITC-dextran as the standard, and the values are expressed as ng/g brain tissue.
33、MX3210-20T Annexin V-FITC/PI Apoptosis Detection Kit 细胞凋亡检测试剂盒
「1」Hou Xy et al.A combination of LightOn gene expression system and tumor microenvironment-responsive nanoparticle delivery system for targeted breast cancer therapy.Acta Pharmaceutica Sinica B Available online 27 April 2020.
Cell-counting-kit 8 (CCK-8), Hoechst 33342, hyaluronidase (HAase) and annexin V-FITC/propidium iodide (PI) apoptosis detection kits were purchased from Shanghai Maokang Biotech Co., Ltd. (Shanghai, China). 4T1 cells (5 × 104 cells/well) were seeded into 6-well plates. After 24 h, the cells were treated with 4 μg of pGDTA (2 μg of pGAVPO and 2 μg of pU5-DTA). After incubation with or without exposure to blue light (2 W/m2) for 24 h, the cells were collected, washed, and stained with annexin V-FITC and propidium iodide (PI). The cells were analyzed for apoptosis using flow cytometry.