1. MCE Kits
  2. Cell Biology
  3. Cell Analysis
  4. Cell Apoptosis and Necrosis
  5. Annexin V-EGFP/PI Apoptosis Detection Kit

Annexin V-EGFP/PI Apoptosis Detection Kit 

Cat. No.: HY-K1074
Manual SDS

Annexin V-EGFP/PI Apoptosis Detection Kit provides a rapid and convenient method to detect cell apoptosis and necrosis. After staining, live cells show little or no fluorescence (Annexin V-/PI-), early apoptosis cells show green fluorescence(Annexin V+/PI-), late apoptosis cells and necrosis cells show red and green fluorescence (Annexin V+/PI+). Compared with FITC, the fluorescence of EGFP is stronger and more stable.


Annexin V-EGFP/PI Apoptosis Detection Kit
Size Price Stock Quantity
20T USD 175 In-stock
50T USD 350 In-stock

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  • Description

  • Storage

  • Protocol

  • Components

  • Documentation

Description
& Advantages

MCE Annexin V-EGFP/PI Apoptosis Detection Kit provides a rapid and convenient method to detect cell apoptosis and necrosis. In normal live cells, Phosphatidylserine (PS) is located on the cytoplasmic surface of the cell membrane. Upon initiation of apoptosis, PS translocates from the inner to the outer leaflet of the membrane. Annexin V is a 35-36 kDa Ca2+ - dependent phospholipid-binding protein that has a high affinity for PS. Annexin V labeled with green fluorescent EGFP can identify apoptotic cells by binding to PS exposed on the outer leaflet. Propidium Iodide (PI) is a cell-membrane impermeable dye to live cells and early apoptosis cells, but stains late apoptosis cells and necrosis cells with red fluorescence. After staining, live cells show little or no fluorescence (Annexin V-/PI-), early apoptosis cells show green fluorescence (Annexin V+/PI-), late apoptosis cells and necrosis cells show red and green fluorescence (Annexin V+/PI+).

Storage

-20°C, 1 year


Protect from light


Avoid repetitive freeze-thaw cycles


Protocol

Incubation of cells with Annexin V-EGFP and PI


1.  Collect 1-5 × 10 cells.


For suspension cells: Centrifuge at 1000 g for 5 minutes and then discard the supernatant. Add 1 mL of pre-cooled PBS to resuspend the cells, centrifuge at 1000 g for 5 minutes and then discard the supernatant.


For adherent cells: Collect the cell culture medium. Wash cells with PBS and add trypsin to dissociate cells. Add the medium and gently suspend the cells to make a single-cell suspension. Centrifuge at 1000 g for 5 minutes and then discard the supernatant. Add 1 mL of pre-cooled PBS to resuspend the cells, centrifuge at 1000 g for 5 minutes and then discard the supernatant.


Note: It is recommended to use trypsin containing no EDTA.


2.  Resuspend the cells in 195 μL of Binding Buffer.


3.  Add 5 μL of Annexin V-EGFP.


4.  Add 10 μL of PI Stain.


5.  Incubate the cells at room temperature for 10-20 minutes in the dark.


Detection by flow cytometer


Analyze Annexin-EGFP binding by flow cytometer (Ex = 488 nm; Em = 507 nm) using EGFP signal detector (usually FL1) and PI staining by the phycoerythrin emission signal detector (usually FL2 or FL3).


Note: It is recommended to perform three controls: a: cells with no Annexin-EGFP or PI; b: cells with only Annexin-EGFP; c: cells with only PI.


Detection by fluorescence microscope


Detect the fluorescence by fluorescence microscope: Centrifuge at 1000 g for 5 minutes, discard the supernatant, resuspend the cells with 50-100 μL of Binding Buffer and then detect the fluorescence by fluorescence microscope.


Components
Components HY-K1074-20T HY-K1074-50T
Annexin V-EGFP 100 μL 250 μL
Binding Buffer 10 mL 26 mL
PI Stain 220 μL 550 μL
Documentation
Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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Product Name:
Annexin V-EGFP/PI Apoptosis Detection Kit
Cat. No.:
HY-K1074
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