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

 
ALX-209-256-T100 100 tests 200.00 USD
 
ALX-209-256-3100 3x100 tests 456.00 USD
Do you need bulk/larger quantities?
 
FITC-labelled recombinant human annexin V fusion protein (produced in E. coli) shows bright green fluorescence (Ex(max): 488nm; Em(max): 530nm).

Product Specification

MW:~52kDa (SDS-PAGE).
 
Source:Produced in E.coli.
 
UniProt ID:P08758
 
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.
 
Concentration:1mg/ml
 
Formulation:Liquid. In PBS, pH 7.4, containing 0.02% sodium azide.
 
Purity:≥90%
 
Purity Detail:Affinity purified.
 
Specificity:Binds to phosphatidylserine (PS).
 
Applications:Flow Cytometry
 
Application Notes:Detection of apoptotic cells by flow cytometry.
 
Shipping:Blue Ice Not Frozen
 
Long Term Storage:+4°C
 
Use/Stability:Stable for at least one year after receipt when stored at +4°C.
 
Handling:Protect from light. Do not freeze.
 
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. 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.
 
209-256
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 competitor’s) 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.
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