Annexin V-FITC

Catalog# BAD1006

Size: 100T

Lot # Check on the product label

Storage Buffer

In PBS (pH7.2), 0.09% NaN₃

Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Application

Flow cytometry (Use 5μl for 10⁵ cells)

It may can work in Immunofluorescence.

 Other applications have not been tested.

The optimal dilutions should be determined by end user.  

Introduction

Annexin A5 (or Annexin V) is a cellular protein in the annexin group. Annexin A5 is used as a probe to detect cells that have expressed phosphatidylserine (PS) on the cell surface, an event found in apoptosis as well as other forms of cell death. The annexin A5 affinity assay typically uses a conjugate of annexin V and a fluorescent or enzymatic label, biotin or other tags, or a radioelement, in a suitable buffer (annexin V binding to PS is Ca2+ dependent). The assay combines annexin V staining of PS membrane events with the staining of cell nucleus with PI or AAD-7 in living cells to distinguish from dead cells.

Staining Protocol

1. Dilute the Binding buffer (10×) to work solution (1×) with distilled water.

2. Harvest cell (about 1×10⁵ cells per test), then wash once with cold PBS, remove the PBS from the cell pellet.

3. Wash again with cold 1× Binding buffer, centrifuge at 300 x g for 10 min, remove the Binding buffer from the cell pellet.

4. Resuspend cells in cold 1× Binding buffer to a concentration of 1×10⁶ cells/mL.

5. Add 100μL of cells (1×10⁵) to each appropriate tube.

6. Add 5μL of Annexin V-FITC to appropriate tubes.

7. Gently vortex each tube and incubate for 10 minutes at room temperature in dark.

8. Add 5μL of PI or 7-AAD solution and incubate for 5 min at room temperature in dark.

9. Wash cells once in PBS and resuspend in PBS.

10. Analyze by flow cytometry within 4 hours.