Protein C is a vitamin K-dependent serine protease that plays a major role in blood coagulation. Binding of Protein C to Endothelial Cell Protein C Receptor (EPCR, also CD201) leads to the proteolytic activation of PAR1 (protease-activated receptor 1) on endothelial cells and subsequent up-regulation of Protein C-induced genes. EPCR is a type I transmembrane glycoprotein in the CD1/MHC family. It is expressed most strongly in endothelial cells of arteries and veins, and in the heart and lungs. Membrane bound EPCR is released by metalloproteolytic cleavage to generate the soluble receptor. The extracellular domain of human and mouse EPCR shares approximately 61% aa sequence homology.
Applications:
Suitable for use in Flow Cytometry. Other applications not tested.
Recommended Dilution:
Flow Cytometry: 10ul labels 1-2.5x10e5 bEnd.3 cells.
Optimal dilutions to be determined by the researcher.
Flow Cytometry Protocol:
1. Fc-block cells with 1ug of mouse IgG/10e5 cells for 15 minutes at RT. Do not wash excess blocking IgG from this reaction.
2. After blocking, add 10ul of P9102-39J to 1-2.5x10e5 cells and incubatedfor 30 minutes at RT.
3. Remove unbound antibody by washing the cells twice in Flow Cytometry Staining Buffer (F4568-10). Note that whole blood requires a RBC lysis step at this point using Flow Cytometry Mouse Lyse Buffer.
4. Resuspend the cells in Flow Cytometry Staining Buffer for final flow cytometric analysis. As a control, cells in a separate tube should be treated with PE-labeled rat IgG2A antibody. This procedure may need to be modified, depending upon cell type and final utilization.
Storage and Stability:
May be stored at 4 degrees C before opening. DO NOT FREEZE! Stable at 4 degrees C as an undiluted liquid. Dilute only prior to immediate use. Stable for at least 12 months at 4 degrees C. Freezing R-Phycoerythrin (PE) conjugates will result in a substantial loss of activity. PE conjugates are sensitive to light.
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