Annexin V-FITC Early Apoptosis Detection Kit (#6592) Datasheet with Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:

IF-IC, FC-L

UniProt ID:

#P08758

Entrez-Gene Id:

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

The components within the Annexin V-FITC Early Apoptosis Staining Kit are stable for 1 year at 4°C. Protect from light.

Specificity / Sensitivity

Species Reactivity:

All Species Expected

Product Description

The Annexin V-FITC Early Apoptosis Detection Kit enables researchers to identify early apoptotic cells within a cell population. Annexin V-FITC conjugated protein binds to cell surfaces expressing phosphatidylserine, an early apotosis marker. Cells stained with propidium iodide (PI), a non-cell-permeable DNA dye, indicate necrotic cells. Cells stained with both PI and annexin V-FITC demonstrate later stage apoptosis and early necrosis. This kit provides enough reagent to perform 100 assays, based on a 250 μl assay volume.

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

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

The Annexin V-FITC conjugate is a product manufactured for Cell Signaling Technology, Inc. by TAU Technologies BV.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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