FITC Annexin V Apoptosis Detection Kit with PI

BioLegend's FITC Annexin V Apoptosis Detection Kit with PI has been specifically designed for the identification of apoptotic and necrotic cells.Annexin V (or Annexin A5) is a member of the annexin family of intracellular proteins that binds to phosphatidylserine (PS) in a calcium-dependent manner. PS is normally only found on the intracellular leaflet of the plasma membrane in healthy cells, but during early apoptosis, membrane asymmetry is lost and PS translocates to the external leaflet. Fluorochrome-labeled Annexin V can then be used to specifically target and identify apoptotic cells. Annexin V Binding Buffer is recommended for use with Annexin V staining. Annexin V binding alone cannot differentiate between apoptotic and necrotic cells. To help distinguish between the necrotic and apoptotic cells we recommend use of our Propidium Iodide Solution (PI). Early apoptotic cells will exclude PI, while late stage apoptotic cells and necrotic cells will stain positively, due to the passage of these dyes into the nucleus where they bind to DNA.Propidium iodide is a fluorescent dye that binds to DNA. When excited by 488 nm laser light, it can be detected with 562-588 nm band pass filter. It is commonly used in evaluation of cell viability or DNA content in cell cycle analysis by flow cytometry.

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SPECIFICATIONS

Catalog Number

640914

Size

100 tests

Applications

FC

Gene Id

308

Dilutions

Staining Procedure: 1. Wash cells twice with cold BioLegend's Cell Staining Buffer, and then resuspend cells in Annexin V Binding Buffer at a concentration of 0.25-1.0 x 107 cells/ml. 2. Transfer 100 microl of cell suspension in a 5 ml test tube. 3. Add 5 microl of FITC Annexin V. 4. Add 10 microl of Propidium Iodide Solution. 5. Gently vortex the cells and incubate for 15 min at room temperature (25C) in the dark. 6. Add 400 microl of Annexin V Binding Buffer to each tube. Analyze by flow cytometry with proper machine settings.

Storage Temp

4 C

Concentration

Shipping Temp

Ambient RT

Storage Buffer

Store at 4C. Do not freeze.Caution: Propidium Iodide Solution is toxigenic and mutagenic; handle with care.

Alternative Names

Kit includes: FITC Annexin V, PI solution, and Binding Buffer

Regulatory Status

RUO