FITC-labelled recombinant human annexin V fusion protein (produced in E. coli) shows bright green fluorescence (Ex(max): 488nm; Em(max): 530nm).
Produced in E.coli.
Sufficient for 100 or 300 tests. 1 test is equivalent to 1μg of the conjugated protein in an experimental volume of 0.4 ml.
Liquid. In PBS, pH 7.4, containing 0.02% sodium azide.
Binds to phosphatidylserine (PS).
Detection of apoptotic cells by flow cytometry.
Blue Ice Not Frozen
Long Term Storage:
Stable for at least one year after receipt when stored at +4°C.
Protect from light. Do not freeze.
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. Incubate 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.
Produced in HEK 293 cells. The extracellular domain of human FasL (APO-1L; CD95L; CD178) (aa 103-281) is fused at the N-terminus to a linker peptide (26 aa) and a FLAG®-tag. Glycosylation of rhsFasL is similar to that of natural human FasL., ≥95% (SDS-PAGE), ELISA | Print as PDF