Apoptosis Assay Kit - FITC

Description

Apoptosis (a programmed cell death) is a well described phenomenon that occurs in many cellular systems. Currently, the most widely used analytical assays are based on monitoring of translocation of phosphatidylserine (PS) from inner phospholipid layer to the cell surface by use of a fluorochrome-labelled Annexin V in combination with appropriate vital dyes.

Apoptosis Assay Kit is based on standard setup that employs Annexin V-FITC conjugate and propidium iodide (PI). The flow cytometry test can discriminate intact cells (FITC-/PI-), apoptotic cells (FITC+/PI-) and necrotic cells (FITC+/PI+).

Format/Formulation

Apoptosis Assay Kit contains following reagents:

1. Annexin V-FITC
Quantity: 0.5 ml. Sufficient for 100 tests.
The reagent is free of unconjugated FITC.
Form: Ready-to-Use solution. No reconstitution is necessary. Reagent is provided in phosphate buffered saline (PBS) containing 15 mM sodium azide and 0.2% (w/v) high-grade protease free Bovine Serum Albumin (BSA) as a stabilizing agent.

2. Binding Buffer
Quantity: 5 ml (10x concentrated).
Form: Concentrated filter-sterilized solution (0.1M HEPES/NaOH, pH 7.4, 1.4M NaCl, 25mM CaCl2). Dilute 10X in deionized water prior use.

3. Propidium Iodide (PI)
Quantity: 0.5 ml.
Form: Ready-to-Use solution in deionized water (0.1 mg/ml).

Related Products

Mouse Monoclonal to CD253 / TRAIL 2E5 (IgG1)

Example Data

JURKAT cells (human T cell leukemia) were treated with 200 ng/ml TRAIL/Apo2L (bacterially expressed extracellular domain of human TRAIL, corresponding to amino acids 95-281) for 6 hours or left untreated.
Detection of apoptotic cells was preformed by flow cytometry using Apoptosis Assay Kit (Cat. No. KIT-ANXF).

Legend:
Green line - untreated and unstained cells
Red line - untreated and stained cells
Blue line - TRAIL-treated and stained cells.

A small proportion of untreated cells were identified as undergoing apoptosis (PS expressing, PI negative). In TRAIL-treated cells the number of early apoptotic cells (PS expressing, PI negative) as well as necrotic cells (PS expressing, PI positive cells) significantly increased.