IF-IC, FC-L
#P08758
308
Product Information
Product Usage Information
1. Dilute the 10X Annexin V Binding Buffer with dH2O to 1X and keep on ice.
2. Collect cells of interest by centrifugation. Wash cells with ice-cold culture medium or PBS.
3. Resuspend cells at 105 to 106 cells/ml with 1X Annexin V Binding Buffer. Aliquot 96 μl cell suspension in an assay tube.
4. Add 1 μl Annexin V-FITC Conjugate and 12.5 μl Propidium Iodide (PI) Solution to each 96 μl cell suspension. Allow cells to incubate 10 min on ice in the dark.
5. Dilute the cell suspension to a final volume of 250 μl/assay with ice-cold, 1X Annexin V Binding Buffer. Analyze immediately..
Storage
Specificity / Sensitivity
Species Reactivity:
All Species Expected
Product Description
Background
Annexin V belongs to the annexin family of proteins consisting of over 160 members. All annexin family members share the same charactersistic of Ca2+-dependent binding to negatively charged phospholipid surfaces (1). During early apoptosis, phosphatidylserine (PS) is translocated from the cytosolic side of the plasma membrane to the cellular surface. This translocation exposes PS to the extracellular environment with the plasma membrane left intact (2). Annexin V protein demonstrates high affinity, specificity, and sensitivity for PS and can be used as a marker of early apoptosis. In order to rule out "leaky" necrotic cells, annexin V must be used in tandem with reagents that determine the integrity of the cell membrane, such as propidium iodide (2).
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Applications Key
IF-IC: Immunofluorescence (Immunocytochemistry) FC-L: Flow Cytometry (Live)
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
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