United Kingdom (UK)
Recombinant monoclonal antibody to Protein C. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma HPC-4.
UniProt Accession Number of Target Protein: P04070
Alternative Name(s) of Target: Autoprothrombin IIA; HPC4; Vitamin K-dependent protein C; Anticoagulant protein C; Blood coagulation factor XIV; PROC
Immunogen: This antibody was raised by immunising mice with human protein C.
Specificity: Specifically binds to a specific twelve peptide sequence (EDQVDPRLIDGK) in the activation region of the Protein C (spanning the thrombin cleavage site). Does not react with activated protein C. Has been shown to be cross-reactive with baboon Protein C but not with bovine or mouse Protein C.
Application Notes: HPC-4 binds human protein C only in the presence of Ca2, it binds to the reduced carboxymethylated protein C heavy chain and it blocks protein C activation but does not bind or inactivate protein C or activated protein C bound to its plasma inhibitor (Taylor et al., 1987). It is an anti-epitope tag antibody, as it recognises EDQVDPRLIDGK sequence (Taylor et al., 1987). Furthermore, it has been shown to work in ELISAs to detemine expression levels (Velander et al., 1992). HPC-4 has been shown to be effective at purifying proteins tagged with its epitope (Rezaie et al., 1992). Has been used to block the activation of Protein C by the thrombin-thrombomodullin complex (Stearns et al., 1988). It was used for the same purpose in Baboons to lower the threshold of sepsis (Taylor Jr et al., 1987). HPC-4 has been used in many other studies to observe the function of Protein C by knocking out its function in a Ca2+ dependent manner, purificaion and others (Bae et al., 2007; Tanaka et al., 2005).
Antibody first published in:
Taylor et al. Protein C prevents the coagulopathic and lethal effects of Escherichia coli infusion in the baboon. J Clin Invest. 1987 Mar;79(3):918-25. PMID:3102560
Note on publication:
Describes the generation and characterisation of the HPC-4 antibody.
ELISA using HPC-4 (Ab01200) and Multi Tag protein 2. Binding curves of the mouse IgG1 chimeric version of the anti-Protein C antibody HPC-4 (Ab01200-1.1; green line) and isotype control (Ab00102-1.1, anti-fluorescein; orange line) to an ELISA plate coated with Multi Tag protein 2 (Pr00565, Absolute Antibody) at a concentration of 5 ug/ml. Ab02007 antibody was titrated on a 3-fold serial dilution starting at 10,000 ng/mL. Anti-mouse IgG was used as secondary antibody for detection at a dilution of 1:4,000.
Western Blot using Anti-Protein C antibody HPC-4 (Ab01200). Protein C expressing plasmid transfected (A) and non-transfected (B) HEK293 cells lysate (3µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric mouse version of HPC-4 (Ab01200-1.1) at 0.001µg/ml before detection using an anti-mouse secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Immunofluorescence staining of HEK293 cells with Anti-Protein C (Ab01200) HPC-4 Immunofluorescence analysis of paraformaldehyde fixed HEK293 cells transfected with Protein C expressing plasmid (A) and non-transfected HEK293 cells (B), permeabilized with 0.15% Triton stained with the chimeric mouse IgG1 version of HPC-4 (Ab01200-1.1) (1:200 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing membrane and cytoplasmic staining. The nuclear stain is DAPI (blue).