Ringer's solution, containing 122 mM sodium chloride, 3 mM potassium chloride, 0.4 mM monopotassium phosphate, 1.2 mM magnesium sulfate, 25 mM sodium bicarbonate, and 1.2 mM calcium chloride, was purchased from Millipore (MA, USA). Pachymic acid, liquiritin, rhynchophylline, isorhynchophylline, corynoxeine, and isocorynoxeine (purity ≥ 99%) were purchased from National Institutes for Food and Drug Control (Beijing, China). Fetal bovine serum (FBS) was purchased from Gibco/BRL (Grand Island, NY, USA). Methyl thiazolyl tetrazolium (MTT), dimethyl sulfoxide (DMSO), and trypsin were provided by Sigma Chemical (St. Louis, MO, USA). Aβ1–42 (purity ≥ 95%) was purchased from Abcam (Cambridge, USA). HPLC-grade acetonitrile was purchased from Thermo Fisher Scientific (Massachusetts, USA). EliVision plus and DAB kits were provided by Fuzhou Maixin Biotech. Co., Ltd. (Fuzhou, China). Bax, Bcl-2, caspase-3 and caspase-9 antibodies, annexin V-FITC/PI assay kit, basal DMEM medium, cell lysis solution, and 0.1% DEPC water were purchased from Nanjing KeyGen Biotech Co., Ltd. (Nanjing, China). Nimodipine was provided by Bayer healthcare Co., Ltd. (Leverkusen, Germany).
Annexin 5 fitc pi assay kit
Manufactured by Keygen Biotech
Sourced in China
The Annexin V-FITC/PI assay kit is a laboratory equipment used to detect and quantify apoptosis, or programmed cell death, in a sample. The kit includes Annexin V conjugated to the fluorescent dye FITC and the DNA-binding dye propidium iodide (PI). The core function of the kit is to differentiate between viable, early apoptotic, and late apoptotic/necrotic cells based on their membrane integrity and phosphatidylserine externalization.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Facscalibur flow cytometer, by BD (2 mentions)
Hplc grade acetonitrile, by Thermo Fisher Scientific (1 mentions)
Aβ1 42, by Abcam (1 mentions)
Trypsin, by Merck Group (1 mentions)
Bcl 2, by Keygen Biotech (1 mentions)
Bcl2 associated x (bax), by Keygen Biotech (1 mentions)
Cell lysis solution, by Keygen Biotech (1 mentions)
Caspase 8 activity assay kit, by Beyotime (1 mentions)
Ac devd cho, by Beyotime (1 mentions)
Caspase 8, by Cell Signaling Technology (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Nf κb p65 8242, by Cell Signaling Technology (1 mentions)
Ao eb, by Beyotime (1 mentions)
Histone h3, by Cell Signaling Technology (1 mentions)
Caspase 9, by Cell Signaling Technology (1 mentions)
Cleaved caspase 8, by Cell Signaling Technology (1 mentions)
Mccoy s 5a medium, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
6 protocols using annexin 5 fitc pi assay kit
1
Neuroprotective Potential of Natural Compounds
2
Assessing Cell Apoptosis by Flow Cytometry
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Cell apoptosis was assessed by flow cytometry using the Annexin V-FITC/PI assay kit (Nanjing KeyGen Biotech, Nanjing, China) to differentiate the survival and apoptotic cells. The HK-2 cells were plated on 6-well plates at initial densities of 5×105 cells/well and incubated for 24 hours prior to the addition of the GNPs. Fresh medium containing GNPs (50 nM) was added to the cells and incubated for another 24 hours. Thereafter, the cells were trypsinized, washed with PBS, resuspended in binding buffer, and incubated with staining solution (annexin V/PI =1:2) in the dark for 20 minutes at room temperature. Immediately after the annexin V/PI staining, fluorescence-activated cell sorting (FACS) analysis was performed using a FACSCalibur™ flow cytometer (BD Biosciences, San Jose, CA, USA).
3
Apoptosis Induction in Cancer Cells
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McCoy's 5A medium, fetal bovine serum (FBS), penicillin-streptomycin and trypsin-EDTA were obtained from Gibco (Grand Island, NY, USA). NAC, Ac-DEVD-CHO, DAPI and AO&EB, DCFH-DA, JC-1 stain kit, Cell and Tissue mitochondria isolation kits, Caspase-8 activity assay kit and BCA protein assay kit were obtained from Beyotime Biotech (Jiangsu, China). Nuclear-Cytosol kit was obtained from Applygen Technologies Inc (Beijing, China). Annexin V-FITC/PI assay kit, ROS and TNF-α ELISA kits were purchased from KeyGEN Biotech (Nanjing, China). Antibodies against Bax (#2772), Bcl-2 (#4223), Cytochrome c (#11940), Caspase-3 (#9662), cleaved Caspase-3 (#9661), Caspase-8 (#4790), cleaved Caspase-8 (#9496), Caspase-9 (#9508), NF-κB (p65, #8242) and Histone H3 (#3638) were purchased from Cell Signalling Technology (Danvers, MA, USA). Anti-β-actin and anti-GAPDH antibodies, anti-mouse and anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibodies, 5-Fluorouracil (5-FU), MTT and standard sugar were purchased from Sigma (St. Louis, MO, USA). All other chemicals used were of analytical grade.
4
Annexin V/PI Apoptosis Assay
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Cell apoptosis was analyzed by Annexin V and propidium iodide (PI) staining using flow cytometry (Peng et al., 2015 (link)), with the Annexin V-FITC/PI assay kit (Nanjing KeyGen Biotech, Nanjing, China) according to the manufacturer’s instructions. FITC Annexin V and PI were 5 μL, respectively, which was added in sequence for cells staining after being trypsinized, centrifuged and resuspended. After incubation with staining solution (annexin V/PI = 1:2) for 20 min in the dark at room temperature. Fluorescence-activated cell sorting (FACS) analysis was immediately performed using a flow cytometer (BD Biosciences, San Jose, CA, USA).
5
Apoptosis Analysis of Cancer Cells
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MDA-MB-231 cells were plated into 6-well culture plates (2 mL/well) and cultured in 5% CO2 at 37 °C overnight. RuDA-NPs or RuDA were added, which were diluted to a concentration of 50 μM. After 12 h incubation, the cells were irradiated with or without 808 nm laser (0.5 W cm−2) for 10 min (300 J cm−2). For the Vitamin C (Vc) group, the cells were treated with 0.5 mM Vc before laser irradiation. After another 12 h, the cells were digested with trypsin and washed twice with cold PBS. Then, cells were collected by centrifugation (450 × g, 5 min). The apoptosis was determined by flow cytometry using an Annexin V-FITC/PI assay kit (KeyGEN BioTECH, China) according to the manufacturer’s protocol. The detailed operation as follows: cells were stained with 5 μL Annexin V-FITC for 5 min in Annexin-binding buffer (10 mM HEPES, 140 mM NaCl, 2.5 mM CaCl2, pH 7.4). PI (propidium iodide) was added to cells with 5 μL before incubating at room temperature for 15 min. The fluorescence of cells was measured by flow cytometer. BD FACSDiva and Cell Quest Pro software were used for data collection and FlowJo V10 was used for data analysis. The results appeared as a percentage of normal and apoptotic cells at various stages.
6
Apoptosis Induction in B16F10 Cells
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B16F10 (3 × 105 cells per well) were seeded in six-well plates and cultured for 24 h. Then, the adherent cells were incubated with PBS, 1MT, GA, G + M, and GM (1MT: 7 μg/ml, GA: 200 μg/ml), respectively, for 48 h. After being washed with cold PBS, they were collected and dyed with Annexin V-FITC/PI assay kit (KeyGEN BioTECH, China).Then, the FACSCalibur flow cytometer (BD Biosciences) was used to analyze cell apoptosis.
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