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Annexin A5 (ANXA5) Protein

Details for Product No. ABIN412008, Supplier: Log in to see
Protein Name
  • anx
  • Anx5
  • ANX5
  • anx5
  • anxa5
  • ANX V
  • cb989
  • ENX2
  • enx2
  • LC5
  • MGC89158
  • PP4
  • R74653
  • RPRGL3
  • wu:fa98f06
  • wu:fj10f10
Fluorescence Microscopy (FM), Flow Cytometry (FACS)
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Purpose Reagent for detecting early stages of apoptosis.
Sample Type Cell Culture
Characteristics The Annexin V-FITC conjugate can be used for detection of apoptosis by fluorescence microscopy or by flow cytometry.
Background During apoptosis, phosphatidylserine (PS) is translocated from the cytoplasmic face of the plasma membrane to the cell surface. Annexin V has a strong, Ca2+-dependent affinity for PS and therefore is used as a probe for detecting apoptosis.
Research Area Cardiovascular, Atherosclerosis, Apoptosis/Necrosis, Coagulation
Pathways Apoptosis
Application Notes Detection method: Flow cytometry (Ex/Em = 488/530 nm) using FL1 channel and fluorescence microscopy
Protocol A. Incubation of cells with Annexin V-FITC:
1. Induce apoptosis by desired methods.
2. Collect 1 x 10^5 cells by centrifugation.
3. Resuspend cells in 500 µL of 1X Annexin V Binding Buffer (ABIN412353).
4. Add 1 µL of Annexin V-FITC and 1 µL Propidium Iodide (ABIN412371).
5. Incubate at room temperature for 5 min in the dark.
Proceed to B or C below depending on method of analysis.

B. Quantification by Flow Cytometry:
Analyze cells by flow cytometry (Ex = 488 nm, Em = 530 nm) using FL1 channel for detecting Annexin v-FITC staining and FL2 channel for detetcing PI staining. For adherent cells, trypsinize and gently wash cells with serum-containing medium before incubation with Annexin V-FITC (A.3-5).

C. Detection by Fluorescence Microscopy:
1. Place the cell suspension from Step A.5 on a glass slide, and cover with a glass coverslip. For analyzing adherent cells, grow cells directly on a coverslip. Following incubation (A.5), invert coverslip on a glass slide and visualize cells. The cells can also be washed with 1X Annexin V Binding Buffer and fixed in 2 % formaldehyebefore visualization. (Cells must be incubated with Annexin V-FITC before fixation because any cell membrane disruption can cause nonspecific binding of annexin V to PS on the inner surface of the cell membrane.)
2. Observe the cells under a fluorescence microscope using a dual filter set for FITC and rhodamine or separate filters. Cells that have bound Annexin V-FITC will show bright green staining on the plasma membrane. Cells which have lost membrane integrity will show red staining (PI) throughout the nucleus and a halo of green staining (FITC) on the cell surface (plasma membrane).

TESTING RESULTS: Jurkat cells (treated with 2 µM camptothecin for 6 hours) were collected for annexin V assay according to the kit instructions. Results show 40-60 % apoptotic cells as analyzed by flow cytometry.
Restrictions For Research Use only
Format Liquid
Concentration 0.15 mg/mL
Buffer PBS containing 1 % BSA, 0.02 % sodium azide
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Do not freeze!
Storage 4 °C
Expiry Date 12 months
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