Cell morphological changes were observed with inverted light microscopy (NIKON, Tokyo, Japan). To identification of the apoptotic induction effect of compounds 6 and 14 , a FITC-labeled Annexin V/PI apoptosis detection Kit (Keygen, Nanjing, China) was used according to the manufacturer’s instructions. Briefly, HL60 cells were exposed to vehicle control (DMSO, <0.1%), compounds 6 (12 μM) and 14 (12 μM). After 48 h, cells were harvested and washed with PBS and resuspended in binding buffer, and then, AnnexinV-FITC and PI were added. After staining for 15 minutes, the cells were immediately analyzed using flow cytometry (Becton Dickinson, USA).
Fitc labeled annexin 5 pi apoptosis detection kit
Manufactured by Keygen Biotech
Sourced in China
The FITC-labeled Annexin V/PI apoptosis detection Kit is a laboratory tool used to detect and quantify apoptosis, a type of programmed cell death. It utilizes Annexin V, a calcium-dependent phospholipid-binding protein, conjugated with the fluorescent dye FITC, and the DNA-binding dye propidium iodide (PI) to differentiate between early apoptotic, late apoptotic, and necrotic cells.
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3 protocols using fitc labeled annexin 5 pi apoptosis detection kit
1
Annexin V Apoptosis Assay of Compounds
2
Apoptosis Induction by Zephycandidine A
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For identification of the apoptotic induction effect of zephycandidine A (1 ), a FITC-labeled Annexin V/PI apoptosis detection Kit (Keygen, Nanjing, China) was used according to the manufacturer’s instructions. Briefly, HL-60 cells were exposure to vehicle control (DMSO, < 0.1%), zephycandidine A (1 ) (3 and 6 μM) for 48 h, VP16 was used as a positive control20 (link). After that cells were harvested and washed with PBS and resuspended in binding buffer, and then, AnnexinV-FITC and PI were added. After staining 15 minutes, the cells were immediately analyzed using flow cytometry (Becton Dickinson, USA).
3
Annexin V/PI Apoptosis Assay
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Apoptosis-mediated cell death of nerve cells was examined using a FITC-labeled Annexin V/PI Apoptosis Detection Kit (KeyGen Biotech, Nanjing, China) according to the manufacturer's instructions. After being harvested and washed with PBS twice, SH-SY5Y and PC12 cells were resuspended in 500 ml binding buffer, followed by adding Annexin V-fluorescein isothiocyanate (5 μl) and PI (5 μl). Then, cells were incubated in the dark for 30 min at room temperature. Flow cytometry (Beckman Coulter, Inc., USA) analysis was done immediately after supravital staining.
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