DSF, CuCl2 solid powder, 3‐(4,5‐dimethylthiazole‐2)‐2,5‐diphenyltetrazolium bromide (MTT) powder 4,4′‐diisothiocyanostibibene‐2,2′‐disulfonic acid (DIDS) powder and 5‐nitro‐2‐ (3‐phenlpropylamino) benzoic acid (NPPB) powder were purchased from Sigma‐Aldrich (St Louis, MO). Annexin V‐FITC/PI apoptosis detection kits were purchased from KeyGEN BioTECH (Jiangsu, China).
Annexin 5 fitc pi apoptosis detection kit
Manufactured by Keygen Biotech
Sourced in China, United States
The Annexin V-FITC/PI apoptosis detection kit is a laboratory reagent used to detect and quantify apoptosis in cell samples. It utilizes Annexin V, a protein that binds to phosphatidylserine, and propidium iodide, a fluorescent dye that binds to DNA, to identify cells undergoing apoptosis.
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Lab products found in correlation
Cell cycle detection kit, by Keygen Biotech (23 mentions)
Facscalibur flow cytometer, by BD (17 mentions)
Facscalibur, by BD (16 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (12 mentions)
Facscan flow cytometer, by BD (11 mentions)
Cellquest software, by BD (10 mentions)
Facscan, by BD (8 mentions)
Facscanto 2, by BD (8 mentions)
Facscanto 2 flow cytometer, by BD (7 mentions)
Flow cytometry, by BD (7 mentions)
Trizol reagent, by Thermo Fisher Scientific (7 mentions)
Cleaved caspase 3, by Cell Signaling Technology (6 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (6 mentions)
Bcl 2, by Cell Signaling Technology (6 mentions)
Accuri c6 flow cytometer, by BD (6 mentions)
Cytoflex, by Beckman Coulter (6 mentions)
Accuri c6, by BD (6 mentions)
β actin, by Cell Signaling Technology (5 mentions)
Novocyte flow cytometer, by Agilent Technologies (5 mentions)
Caspase 3, by Cell Signaling Technology (5 mentions)
330 protocols using annexin 5 fitc pi apoptosis detection kit
1
Apoptosis Detection in Cell Lines
2
Evaluation of RY10-4 Anti-cancer Effects
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RY10-4 (>95%) was prepared previously in our laboratory as described by Yuan et al. [5 (link)]. Sulforhodamine B (SRB) was purchased from J&K Scientific (Beijing, China). Dimethyl sulfoxide (DMSO) and N-acetylcysteine (NAC) were purchased from Sigma Aldrich (St. Louis, MO, USA). Hoechst 33342, propidium iodide (PI), reactive oxygen species assay kit (DCFH-DA), RIPA lysis buffer, BCA protein assay kit, and BeyoECL plus chemiluminescence kit were obtained from Beyotime Inc. (Shanghai, China). Annexin V-FITC/PI apoptosis detection kits was purchased from KeyGEN BioTECH (Nanjing, China). Specific antibodies against Bcl-2, Bax, p53, cyclin E, CDK2, cleaved caspase3, actin, STAT3, p-STAT3, p21, GAPDH and corresponding secondary antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA).
3
Guaiol-Induced Apoptosis Pathway
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(−)-Guaiol was commercially obtained from Sigma (448575, St. Louis, MO, USA). DMEM medium was purchased from KeyGEN BioTECH (KGM12800-500, Jiangsu, China). Fetal bovine serum (FBS) was provided by Gibco Life Technologies (10099-141, Grand Island, NY, USA). The reagent Z-VAD-FMK (Z-VAD) was obtained from Selleck (S7023, Shanghai, China). Annexin V-FITC/PI Apoptosis Detection Kits were purchased from KeyGEN BioTECH (KGA105-KGA108, Jiangsu, China). Cell Counting Kit-8 was purchased from YEASEN (40203ES76, Shanghai, China). LY294002 was purchased from MedChemExpress (HY-10108, Monmouth Junction, New Jersey, USA). SYTOX Green was purchased from Invitrogen (S7020, Carlsbad, California, USA). The following antibodies – Bcl-2(15071, 1: 1000), BAX (5023, 1: 1000), β-actin (4970, 1: 1000), p-Akt (4060, 1: 1000), Akt (2920, 1: 1000), PI3K (4255, 1: 1000), p-PI3K (4249, 1: 1000), and IgG secondary antibody (10700, 1: 2000) – were purchased from Cell Signaling Technology (Boston, Massachusetts, USA).
4
Annexin V-FITC/PI Apoptosis Assay
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Apoptosis assay was carried out using the Annexin VFITC/PI apoptosis detection kit (KeyGen Biotech) according to the manufacturer's protocol. Briefly, 0.5 mL binding buffer was added into 1 × 105 A549 and HBE cells. Sequentially, the cells were stained with PI at room temperature for 15 min and then analysed by flow cytometry (BD, FACSCantoTM II).
5
Annexin V-FITC/PI Apoptosis Assay
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Cell apoptosis was measured by flow cytometry following the instructions of the Annexin V-FITC/PI apoptosis detection kit (Nanjing KeyGen Biotech Co., Ltd., Nanjing, China). Cultured cells were harvested and washed twice in PBS, re-suspended in binding buffer, and then incubated with Annexin V-FITC and propidium iodide (PI) for 15 min at room temperature. Afterwards, flow cytometry was performed to determine rate of apoptosis on a FACSAria flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA).
6
Annexin V-FITC/PI Apoptosis Assay
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Cell apoptosis was evaluated using the Annexin V-FITC/PI apoptosis detection kit (Keygen Biotech, Nanjing, China) according to the manufacturer’s instructions. Briefly, cells treated with or without 3.82 or 2.85 mg/mL MI were seeded into 6-well plates at a density of approximately 1 × 105 cells/mL. The cells were collected, harvested, washed, and re-suspended in the binding buffer. After stained with Annexin V-FITC at room temperature for 20 min in the dark, cells were incubated with PI for another 10 min. The apoptosis rate of cells was analyzed using a FACSCantonTMII flow cytometer (FACSCantonTMII, BD, Shanghai, China).
7
Apoptosis Modulation in Jurkat Cells
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The cell co-culture system was established according to the above methods. Jurkat cells without activation of PHA were used as control. Changes in apoptosis level were investigated in Jurkat cells activated by 2 μg/mL PHA for 48 h, co-cultured for 48 h and co-cultured for 48 h with PD-1 mAb B1C4 (5 μg/mL) added. The effect of co-culture system and PD-1 mAb B1C4 on apoptosis of Jurkat cells was measured. Morphological characteristics of chromatin indicating cell apoptosis were observed by DAPI staining under a confocal microscope (Leica). The early and late apoptosis cells were quantitatively estimated by Annexin V-FITC/PI double staining from the NovoCyte flow cytometer (ACEA Biosciences) using an Annexin V-FITC/PI apoptosis detection kit (KeyGEN BioTECH).
8
Annexin V-FITC/PI Apoptosis Assay
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Apoptosis rates were assessed using the Annexin V-FITC/PI Apoptosis Detection Kit (KeyGEN BioTECH, Nanjing, China) according to the instructions from the manufacturer. The cells were seeded into 6-well tissue culture plates (4 × 10 (5 (link)) cells/well). Following treatment, the cells were collected, washed with PBS, and resuspended in 500 μL binding buffer. Then, 5 μL Annexin V-FITC and 5 μL PI were added to the buffer and incubated at room temperature for 15 min in the dark. Cells were analyzed by flow cytometry (BD FACSCanto) within 1 hour.
9
Annexin V-FITC/PI Apoptosis Assay
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Apoptotic cell numbers were detected by Annexin V-FITC/PI Apoptosis Detection Kit (Nanjing KeyGen Biotech Co. Ltd., Nanjing, China). Twenty-four hours after different treatment, cells were digested by trypsin without EDTA, and resuscitated at a concentration of 1× 10 6 cells /mL in 1× binding buffer. Annexin V-FITC and PI were added into the binding buffer and well mixed. After 30 minutes incubation with room tempreture without light, the ow cytometry (BD Biosciences, San Diego, CA) was used to detect the signals of cells.
10
Apoptosis and Cell Cycle Analysis
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Cells (5 × 104 cells/well) were seeded into six-well plates. The medium was replaced with 2 ml of experimental medium for 24 h, followed by serum-free experimental medium for an additional 24 h. After the cells were harvested, the rate of apoptosis was determined using an Annexin V-FITC/PI apoptosis detection kit (KeyGen Biotechnology Co., Ltd., Nanjing, China) according to the manufacturer’s instructions. Data analysis was performed using FlowJo software. After 48 h, the cells were collected and fixed in 70% ethanol overnight at 4 °C then stained with propidium iodide. A FACScan flow cytometer (Biosciences, San Jose, CA, USA) was utilized for cell cycle analysis. Data was analyzed using ModFit 3.0 software (Verity Software House, Topsham, ME, USA).
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