Annexin A5 (ANXA5) Protein

Details for Product No. ABIN412007
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Protein Name
Synonyms anx, anx5, ANX V, anxa5, cb989, wu:fa98f06, wu:fj10f10, MGC89158, ANX5, ENX2, PP4, RPRGL3, Anx5, R74653, LC5, enx2
(36), (4), (4), (4), (4)
(34), (5), (2), (2), (2)
Flow Cytometry (FACS), Fluorescence Microscopy (FM)
Pubmed 56 references available
Quantity 1000 tests
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Catalog No. ABIN412007
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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.
Alternative Name Annexin V
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
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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