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CellSimple™ AnnexinV-FITC Early Apoptosis Detection Kit

  1. Dilute the 10X Annexin V Binding buffer with 1X dH2O 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 - 106 cells / ml with 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, using the Apoptosis (Annexin V-FITC, PI) App on the CellSimple™ Cell Analyzer.

posted April 2016

Product Includes Quantity (with Count)
Annexin V-FITC Conjugate 1 x 100 µl
10X Annexin V Binding Buffer 2 x 1.5 ml
Propidium Iodide (PI) Solution 1 x 1.3 ml

Product Description

The CellSimple™ Annexin V-FITC Early Apoptosis Detection Kit is designed for use with the CellSimple™ Cell Analyzer. It allows 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. Necrotic cells can be identified by staining with propidium iodide (PI), a non-cell-permeable DNA dye. Cells stained with both PI and annexin V-FITC are indicative of later stage apoptosis and early necrosis. This kit can be used together with the Apoptosis (Annexin V-FITC, PI) App on the CellSimple™ Cell Analyzer. This kit provides enough reagent to perform 100 assays, based on a 250 μl assay volume. The CellSimple™ Cell Analyzer comes complete with a pack of 25 cassettes (50 tests), a USB cord and power adapter, and user documentation.  

The CellSimple™ Cell Analyzer is currently only available in the US.

Product Usage Information

Storage: Store at 4°C. Protect from light. Kit components are stable for 12 months.

Specificity / Sensitivity

Species Reactivity: Human
Species predicted to react based on 100% sequence homology: All Species Expected

The CellSimple™ Cell Analyzer is a benchtop instrument that utilizes a disposable thin-film cassette and a combination of a 488 nm laser, two photomultiplier tubes (525/45 nm and 561 nm LP filters), Coulter Principle-based cell measurements, and on-board software to provide easy-to-run applications and data analysis. Data acquisition occurs within approximately 10 seconds per test. The instrument relies on disposable cassettes for sample handling, which alleviates the need for flow cell cleaning and fluidics maintenance and the instrument is small enough to be portable between the lab bench and the hood. Applications include quantitative assessments of cell viability, apoptosis, other labeled antibody markers and single and multiplexed bead-based assays for protein and cellular analysis.

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).

1.  Boersma, H.H. et al. (2005) J Nucl Med 46, 2035-50.

2.  Vermes, I. et al. (1995) J Immunol Methods 184, 39-51.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
CellSimple is a trademark of Cell Signaling Technology, Inc.

CellSimple™ Annexin V-FITC Early Apoptosis Detection Kit