Uses
Anti-Protein C Affinity Matrix is used for:
- Immunoprecipitation of Protein C-tagged proteins from mammalian, bacterial, and yeast cell extracts
- Affinity column purification of Protein C-tagged proteins from crude protein extracts
Following immunoprecipitation or purification, the tagged protein of interest may be analyzed by:
- Western blotting using the Anti-Protein C antibody
- Silver staining (or similar protein stain)
General Description
Protein C is a Vitamin K-dependent plasma zymogen that is activated by proteolytic cleavage of the thrombin-thrombomodulin complex to form an anticoagulant enzyme. Anti-Protein C mouse monoclonal antibody (clone HPC4) binds specifically to an epitope sequence spanning the thrombin cleavage site of protein C and is immobilized. Anti-Protein C recognizes the 12-amino acid sequence (EDQVDPRLIDGK), which encodes residues 6 through 17 of the heavy chain of Protein C. The formation of the Anti-Protein C/protein C epitope complex is dependent on the presence of calcium ions. In the presence of Ca
2+, the antibody binds with high affinity and specificity to this sequence in native human Protein C or in proteins tagged with this epitope. This efficient binding within the recombinant fusion protein occurs regardless of the site of incorporation of the epitope tag (i.e., N terminus, C terminus, or within the reading frame). This unique antibody is especially well suited for purification of recombinant fusion proteins tagged with the protein C epitope.
- Insertion of the protein C tag does not introduce a new metal-ion binding site. The antibody contains the Ca2+ binding site.
- Protein C tag can be integrated either at the N-terminus, C-terminus or internally without any change in antibody specificity.
- Rapid immunoaffinity purification under non-denaturing conditions using economical calcium chelating agent (e.g., EDTA) or alternatively a specific protein C-tag peptide.
Monoclonal mouse antibody Anti-Protein C (clone HPC4) is covalently coupled to agarose beads. In the coupling reaction 4mg of antibody is reacted per 1ml of beads.
