FITC-labelled recombinant human annexin V fusion protein (produced in E. coli) shows bright green fluorescence (Ex(max): 488nm; Em(max): 530nm).
Product Details
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.
Regulatory Status:
RUO - Research Use Only
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
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Product Literature References
Dopamine induces apoptosis in APPswe-expressing Neuro2A cells following Pepstatin-sensitive proteolysis of APP in acid compartments: M. Cagnin, et al.; Brain Res. 1471, 102 (2012), Abstract;
Upregulation of RGS4 and downregulation of CPI-17 mediate inhibition of colonic muscle contraction by interleukin-1beta: W. Hu, et al.; Am. J. Physiol. Cell Physiol. 293, 1991 (2007), Abstract; Full Text
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