Apoptosis Detection Kit (Annexin-V-FITC / PI)

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Antigen
  • anx
  • anx5
  • ANX V
  • anxa5
  • cb989
  • wu:fa98f06
  • wu:fj10f10
  • MGC89158
  • ANX5
  • ENX2
  • PP4
  • RPRGL3
  • Anx5
  • R74653
  • LC5
  • enx2
  • annexin A5
  • annexin A5b
  • Annexin A5
  • annexin 5
  • annexin A5-like
  • ANXA5
  • anxa5b
  • anxa5
  • LOC100220003
  • LOC100305020
  • LOC100342904
  • Anxa5
  • LOC100521982
Alternatives
Sheep (Ovine) Annexin V ELISA Kit
Conjugate
FITC
Application
Flow Cytometry (FACS)
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Purpose This Kit is designed to rapidly analyse different stages of apoptosis and cell death in single cell suspensions.
Characteristics Annexin V binds specifically to phosphatidylserine (PS) which is located to the inner side of the cytoplasmic membrane in intact cells. But after initiation of apoptosis PS becomes rapidly translocated to the outer side of the cytoplasmic membrane where it is thought to play an important role in macrophage recognition, thus allowing the apoptotic cells to become rapidly phagocytosed. The binding of annexin V to PS is Ca2+ -dependent and therefore, a specific Ca2+ -containing reaction buffer is needed during the binding process. At low PS concentrations a binding ratio of eight annexin V molecules to one PS has been reported making annexin V- conjugates ideal for identifying membrane changes associated with early apoptotic events. In late apoptosis/necrosis cells loose their membrane integrity which allows further annexin V binding to PS located to the inner cell membrane. Therefore, MabTag's Annexin V Apoptosis Detection Kit contains a viability dye such as propidium iodide (PI). This DNAintercalating dye can only pass membranes which lost their integrity and therefore, allow the discrimination of early apoptotic cells (Annexin V+ /PI-) from late apoptotic/necrotic cells (Annexin V+ /PI+).
Components 500 μl Annexin V FITC conjugate, (with 1% BSA / 0.1% NaN3 as preservative)
500 μl propidium iodide [20 μg/ml], (w/o preservative)
10 ml Annexin V binding buffer (10fold concentrate), (w/o preservative)
Material not included
  • Flow cytometer or fluorescence microscope
  • Microcentrifuge
  • Microcentrifuge tubes
  • Adjustable pipettors
  • Pipette tips
  • Gloves
  • Deionized or destilled water
  • PBS and/or culture medium
Alternative Name Annexin-V (ANXA5 ELISA Kit Abstract)
Assay Procedure
  1. Dilute an appropriate amount of 10fold Annexin V binding buffer with deionized or
    distilled water in 1:10 ratio (mix 1 part 10x buffer with 9 parts water)
    2. Wash cells with culture medium or PBS
    3. Resuspend 104 - 106 cells in 90 μL of diluted (1 fold) Annexin V binding buffer.
    4. Add 5 μL of Annexin V conjugate and 5 μL of propidium iodide solution.
    5. Incubate for 20 minutes in the dark.
    6. Add 400 μL of Annexin V binding buffer (1fold).
    7. Centrifuge at 400 x g for 5 minutes.
    8. Resuspend cells in 200 μL Annexin V binding buffer (1fold) and analyze sample by flow cytometry or fluorescence microscopy.
Restrictions For Research Use only
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage 4 °C
Storage Comment Upon arrival store all reagents in the dark at 2 - 8 °C. If stored properly all reagents will be stable for 12 months
Expiry Date 12 months
Supplier Images
Flow Cytometry (FACS) image for Apoptosis Detection Kit (Annexin-V-FITC / PI) (ABIN2746064) 1. viable cells (double-negative)
2. early apoptotic cells (annexin V-positive...
Product cited in: Honisch, Yu, Liu, Alesutan, Towhid, Tsapara, Schleicher, Handgretinger, Stournaras, Lang: "Chorein addiction in VPS13A overexpressing rhabdomyosarcoma cells." in: Oncotarget, Vol. 6, Issue 12, pp. 10309-19, 2015 (PubMed).