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Annexin V-FITC/PI Apoptosis Test Kit [BY-L-10012]

Cat. No.: BY-L-10012
Size: 20T/50T/100T
Application: Detecting early apoptosis by flow cytometry or fluorescence microscope
Feature: Low background, high sensitivity
Storage: Keep in dark place, Store at 4℃ for 12 months
Product Detail

Product Introduction
Cell apoptosis is one of the basic characteristics of cells, which plays a very important role in embryonic development, tissue repair, and the stability of the internal environment of the body. Annexin V is a 35.8 KD calcium-dependent phospholipid-binding protein that can specifically bind with high affinity to phosphatidylserine (PS) that is translocated from inner to outer membrane during cell apoptosis. FITC-Annexin V binds to apoptotic cells and emits green fluorescence under blue light excitation, distinguishing apoptotic cells from normal cells. Propidium iodide (PI) is a nucleic acid dye that cannot penetrate intact cell membranes, but can penetrate damaged cell membranes of late-stage apoptotic cells and dead cells, staining the nuclei of cells red. Co-staining FITC-Annexin V with PI can distinguish early apoptotic cells from late apoptotic cells.


Flow Cytometry Analysis
FITC has a maximum excitation wavelength of 488 nm and a maximum emission wavelength of 525 nm. The green fluorescence of FITC is detected in the FL1 channel, while the PI-DNA complex has a maximum excitation wavelength of 535 nm and a maximum emission wavelength of 615 nm. The red fluorescence of PI is detected in the FL2 or FL3 channel. Analysis can be performed using software such as FlowJo and CellQuest, and a dual-color dot plot can be drawn with FITC as the x-axis and PI as the y-axis. In typical experiments, cells can be divided into three subsets: live cells with only a low-intensity background fluorescence, early apoptotic cells with only strong green fluorescence, and late apoptotic cells with both green and red fluorescence staining.

Fluorescence Microscope Analysis

1.Drop a droplet of cell suspension stained with Annexin V-FITC/PI onto a microscope slide and cover the cells with a coverslip. Note: for adherent cells, the cells can be directly cultured on the coverslip and induced to undergo apoptosis.

2.Observe under a fluorescence microscope using dual-color filters. The Annexin V-FITC fluorescence signal appears green and the PI fluorescence signal appears red.

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