Annexin V (human), (recombinant) (FITC conjugate)



ALX-209-256-T100	100 tests	175.00 USD

ALX-209-256-3100	3x100 tests	385.00 USD
Bulk Request

FITC-labelled recombinant human annexin V fusion protein (produced in E. coli) shows bright green fluorescence (Ex(max): 488nm; Em(max): 530nm).

Product Specification

Concentration:	1mg/ml

Purity Detail:	Affinity purified.

MW:	~52kDa (SDS-PAGE).

Quantity:	Sufficient for 100 or 300 tests. 1 test is equivalent to 1μg of the conjugated protein in an experimental volume of 0.4 ml.

Purity:	≥90%

Formulation:	Liquid. In PBS, pH 7.4, containing 0.02% sodium azide.

Specificity:	Binds to phosphatidylserine (PS).

Application:	Detection of apoptotic cells by flow cytometry.

Long Term Storage:	+4°C

Use/Stability:	Stable for at least one year after receipt when stored at +4°C.

Handling:	Do not freeze. Protect from light.

Background / Technical Information:	UniProt ID P08758: Annexin V (human) AfCS Signaling Gateway ID A000281: Annexin V (mouse)

Protocol:	Flow Cytometry Protocol: -Treat cells for induction of apoptosis; As a recommended positive control, incubate with FasL (Prod. No. ALX-522-001) for 6 hours. -Wash cells in PBS. -Resuspend 200,000 cells in 0.4ml binding buffer (10mM HEPES/NaOH, pH7.4, 140mM sodium cloride, 2.5mM calcium cloride, filtered with 0.2μm filter. - Add 1μl Annexin V-FITC to a final concentration of 2.5μg/ml. Incubqate for 10 minutes in the dark. -Wash cells once in binding buffer and resuspend in 0.4ml binding buffer. - Analyze by FACS. Note: These are general recommendations. Optimal conditions must be determined individually for each application.

Figure A: FACS histogram for the fluorescence intensity measured in FL-1 (FITC) channel.
Jurkat cells were incubated for 2 or 6 hours with medium (untreated) or with 100ng/ml rhsFasL in the presence of 1µg/ml Enhancer for Ligands (rhsFasL Set, Prod. No. ALX-850-014).

Figure B: Comparison of ALEXIS and Competitor Annexin-V. Jurkat cells were incubated for 2 or 6 hours with medium (background) or with 100ng/ml rhs FasL in the presence of 1µg/ml Enhancer for Ligands (rhs FasL Set, Prod. No. 850-014). Cells were stained with Annexin-V-FITC (Prod. No. 209-256 and competitors) for 10 min. and then analyzed on FACS. Shown here is the increase in Annexin-V-FITC positive cells as increase from the background. Same amounts of Annexin-V-FITC were compared.

Method: Jurkat cells were incubated for 2 or 6 hours with medium (background) or with 100ng/ml rhsFasL in the presence of 1µg/ml Enhancer for Ligands (rhsFasL Set, Prod. No. ALX-850-014). 2x10⁵ cells per staining were washed once in PBS, resuspended in binding buffer and stained with 5µl Annexin-V-FITC (Prod. No. ALX-209-256 and from competitor) for 10 min. at RT in the dark. Cells were then washed once, resuspended in binding buffer, and analyzed by FACS.

Please mouse over