MC3T3 E1 and RAW264.7 were seeded for 24 h, and then the medium was replaced with serum-free medium (BLANK), control CCM, and RUNX2 k/in CCM. After 24 h of incubation, cell viability was detected by the CCK-8 assay (Vazyme: A311-01), apoptosis was detected with the Annexin V-APC/PI Apoptosis Kit (KeyGEN Biotech: KGA1030), and differentiation was detected by quantitative PCR according to the manufacturer’s instructions.
Annexin 5 apc pi apoptosis kit
Manufactured by Keygen Biotech
Sourced in China
The Annexin V-APC/PI Apoptosis Kit is a laboratory tool used to detect and quantify apoptosis, a form of programmed cell death. The kit combines Annexin V, a protein that binds to phosphatidylserine, and propidium iodide (PI), a DNA-binding dye. This combination allows for the identification of early and late apoptotic cells through flow cytometry analysis.
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Lab products found in correlation
Cck 8 assay, by Vazyme (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Pd 1 pe cy7, by Thermo Fisher Scientific (1 mentions)
Cd3 apc, by Thermo Fisher Scientific (1 mentions)
Cd8 pe, by Thermo Fisher Scientific (1 mentions)
Cd44 pe cy7, by Thermo Fisher Scientific (1 mentions)
Perforin apc, by BioLegend (1 mentions)
Cd4 fitc, by Thermo Fisher Scientific (1 mentions)
Facscanto 2, by BD (1 mentions)
4 protocols using annexin 5 apc pi apoptosis kit
1
Cell Viability, Apoptosis, and Differentiation
2
Annexin V-APC/PI Apoptosis Assay
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Cell apoptosis was determined by Annexin V-APC/PI apoptosis kit (Keygen Biotech, Nanjing, China), followed by flow cytometer analysis (BD FACSCalibur flow cytometer, Becton & Dickinson Company, Franklin Lakes, NJ, USA) according to the manufacturer’s instructions. The percentages of apoptotic cells in the Annexin V+/PI− and Annexin V+/PI+ populations were determined.
3
Assessing Apoptosis in Cancer Cell Lines
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Activated T cells were co-cultured with SNU-449 or SK-Hep-1 at different E/T ratios for 24 h. The apoptosis percentage of SNU-449 and SK-Hep-1 cells was detected using flow cytometry using Annexin V-APC/PI Apoptosis Kit (KeyGEN BioTECH) according to the manufacturer’s instructions.
For flow cytometry, cells were washed in PBS and centrifuged at an appropriate speed. The cells were then fixed with 4% PFA, washed twice, and blocked with 5% BSA in PBS. After centrifugation, the cells were incubated in the primary antibodies for 30 min. The cells were then washed again and analyzed using flow cytometry (Becton Dickinson). For each independent sample, a total of 10,000 cells were collected for analysis.
The primary antibodies were as follows: GzmB-PE (372208; BioLegend), PD-L1-APC (17-5938-42; Invitrogen), perforin-APC (308112; BioLegend), CD62L-PC5.5 (45-0621-82; eBioscience), CD8-PE (12-0084-82; eBioscience), PD-1-PECy7 (25-9985-82; eBioscience), CD4-FITC (11-0041-82; eBioscience), CD3-APC (17-0031-82; eBioscience), CD8-PEcp-cy5.5 (45-0088-42; eBioscience), and CD44-PECy7 (25-0441-82; eBioscience).
For flow cytometry, cells were washed in PBS and centrifuged at an appropriate speed. The cells were then fixed with 4% PFA, washed twice, and blocked with 5% BSA in PBS. After centrifugation, the cells were incubated in the primary antibodies for 30 min. The cells were then washed again and analyzed using flow cytometry (Becton Dickinson). For each independent sample, a total of 10,000 cells were collected for analysis.
The primary antibodies were as follows: GzmB-PE (372208; BioLegend), PD-L1-APC (17-5938-42; Invitrogen), perforin-APC (308112; BioLegend), CD62L-PC5.5 (45-0621-82; eBioscience), CD8-PE (12-0084-82; eBioscience), PD-1-PECy7 (25-9985-82; eBioscience), CD4-FITC (11-0041-82; eBioscience), CD3-APC (17-0031-82; eBioscience), CD8-PEcp-cy5.5 (45-0088-42; eBioscience), and CD44-PECy7 (25-0441-82; eBioscience).
4
Annexin V-APC/PI Apoptosis Assay
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Cell apoptosis was determined by Annexin V-APC/PI apoptosis kit (Keygen Biotech, Nanjing, China), followed by flow cytometer analysis (BD FACSCanto II, BD biosciences) according to manufacturer's instructions. The percentages of apoptotic cells in the Annexin V+/PI− and Annexin V+/PI+ populations were determined.
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