Apoptosis assay in cells was performed using an Annexin V-PE/7-AAD Apoptosis Assay Kit (KeyGEN BioTECH) according to manufacturer’s instructions. Briefly, A498 cells were harvested and incubated with Annexin V-PE and 7-AAD for 15 min. Apoptotic cells populations were analyzed by FACS system. The cell cycle phase distribution were determined by propidium iodide (KeyGEN BioTECH) staining. Briefly, A498 cells were harvested and and fixed with 70% ice-cold ethanol for at least 2 h. After centrifugation at 1000 rpm for 5 min, the cells were washed and resuspended with 0.5 mL PI/RNase buffer for 30 min in the dark. Subsequently, the cell cycle distribution was tested by FACS system.
Annexin 5 pe 7 aad apoptosis assay kit
Manufactured by Keygen Biotech
Sourced in China
The Annexin V-PE/7-AAD Apoptosis Assay Kit is a laboratory tool used to detect and quantify apoptosis, a type of programmed cell death. The kit utilizes the fluorescent dyes Annexin V-PE and 7-AAD to identify cells undergoing early and late stages of apoptosis, respectively. This allows for the differentiation between viable, early apoptotic, and late apoptotic/necrotic cells.
Automatically generated - may contain errors
Lab products found in correlation
Propidium iodide, by Keygen Biotech (1 mentions)
Cell counting kit 8 cck 8, by Beyotime (1 mentions)
Caspase 9 activity assay kit, by Beyotime (1 mentions)
Epirubicin, by Pfizer (1 mentions)
Lactate dehydrogenase ldh cytotoxicity assay kit, by Beyotime (1 mentions)
Anti gapdh antibody, by MultiSciences Biotech (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Facscalibur flow cytometry system, by BD (1 mentions)
4 protocols using annexin 5 pe 7 aad apoptosis assay kit
1
Apoptosis and Cell Cycle Analysis
2
Autophagy Modulation by Epirubicin and BAF1
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Epirubicin was from Pfizer Pharmaceuticals (Dalian, China). Bafilomycin A1 (BAF1) was from Sigma‐Aldrich (Shanghai) Trading (Shanghai, China). The annexin V‐PE/7‐AAD Apoptosis Assay Kit was from Nanjing KeyGen Biotech (Nanjing, China). The caspase‐9 activity assay kit, the lactate dehydrogenase (LDH) cytotoxicity assay kit, and the Cell Counting Kit‐8 (CCK‐8) were from the Beyotime Institute of Biotechnology (Shanghai, China). Anti‐MAPLC3β (H50) and anti‐Ambra1 antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti‐GAPDH antibody was from MultiSciences (Lianke) Biotech (Shanghai, China). Anti‐p62, ATG12, and ATG5 were from Abcam (HongKong, China). Anti‐Beclin 1 was from CST (Danvers, MA, USA).
3
Gemcitabine-Induced Apoptosis Assay
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We harvested cells in the exponential growth phase by incubating them with trypsin without EDTA and seeded them into 6-well plates to detect apoptosis. Once the cells adhered to the plate, the experimental group was treated with gemcitabine (0.01 μg/ml for CAPAN-1 cells and 50 μg/ml for PANC-1 cells). The cells were then cultured for 72 h, and an Annexin V-PE/7-AAD Apoptosis Assay Kit (KeyGEN BioTECH) was subsequently used to measure the percentage of apoptotic cells. Apoptosis was analyzed using a BD FACSCalibur flow cytometer within 1 h.
4
Quantifying HT-29 Cell Apoptosis
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The apoptosis of HT-29 cells was detected by flow cytometry using an Annexin V-PE/7-AAD apoptosis assay kit (Nanjing KeyGen Biotech Co., Ltd., Nanjing, China). Briefly, HT-29 cells were planted into 6-well plates. After culturing for 24 h, cells were incubated with 2.5 μM 5-Fu for 48 h. Cells were collected and washed twice with PBS. Then, cells were incubated 15 min in the dark at room temperature with 5 μL of 7-AAD solution, which was dissolved in 50 μL of Binding Buffer. After adding 450 μl of Binding Buffer, cells were maintained with Annexin V-PE (1 μL) for 15 min in the dark. Finally, cells were measured by a BD FACS Calibur Flow Cytometry System (BD Biosciences, Franklin Lakes, NJ, USA). Annexin V-PE-positive cells were regarded as apoptotic cells.
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