Annexin V-FITC apoptosis detection kit

FITC-labelled recombinant human annexin V shows bright green fluorescence (Ex(max): 488nm, Em(max): 530nm).

Purity:	≥98% (SDS-PAGE; HPLC)

Kit/Set Contains:	Annexin V-FITC: 100 tests/500µl; 300 tests/1.5ml;Liquid. In 50mM TRIS, pH 7.4, 100mM sodium chloride, 1% BSA and 0.02% sodium azide. Binding Buffer: 50ml (4x concentrated)Concentration after dilution: 10mM HEPES/NaOH, pH 7.4, 140mM sodium chloride, 2.5mM CaCl_2. Propidium Iodide: 20µg/ml (100 tests: 1.6ml, 300 tests: 2 x 1.6ml).

Application:	For the detection of apoptotic cells by flow cytometry.

Long Term Storage:	+4°C

Handling:	Avoid freeze/thaw cycles. After opening annexin V-FITC, aliquot contents and freeze at -20°C.

Protocol:	Flow Cytometry Protocol: - Dilute binding buffer 1:4 in distilled water (50ml binding buffer and 150ml water) - Brief centrifugation of annexin V-FITC before use is recommended - Wash cells in PBS by gentle shaking or pipetting up and down. - Resuspend cells in prediluted binding buffer; adjust cell density to 2-5x10⁵/ml - Take 195µl cell suspension and add 5µl annexin V-FITC - Mix and incubate 10 minutes in the dark at room temperature - Wash cells 1x with PBS and resuspend in 190µl prediluted binding buffer - Add 10µl of 20µg/ml propidium iodide stock solution (end concentration 1µg/ml) - FACS analysis