Anti-Proteinase 3/PR3 Antibody, clone MCPR3-3

Myeloblastin (EC 3.4.21.76; UniProt P24158; also known as AGP7, C-ANCA antigen, Leukocyte proteinase 3, Neutrophil proteinase 4, NP-4, P29, PR-3, PR3, Wegener autoantigen) is encoded by the PRTN3 (also known as CANCA, MBN, NP4) gene (Gene ID 5657) in human. Proteinase 3 (PR3) is one of four neutral serine proteases (elastase, cathepsin G, PR3, and neutrophil serine protease 4) stored as fully processed mature enzymes in azurophil granules of human neutrophils. Instead of being targeted to granules after their synthesis, some PR3 molecules end up on the surface of neutrophil plasma membrane in either pro- or mature form. The degree of such surface expression is genetically determined, but the surface exposure and pericellular activity of PR3 around neutrophils can be further upregulated upon neutrophil priming and activation. PR3 autoantibody (anti-neutrophil cytoplasmic antibodies or ANCA) is the main pathogenic feature in patients suffering from granulomatosis with polyangiitis (GPA; formerly called Wegener granulomatosis). ANCAs are shown to activate cytokine-primed neutrophils in vitro by cross-linking surface-exposed PR3 and Fcγ receptors. PR3 activity and/or its inactivation by alpha 1-antitrypsin/alpha-1 proteinase inhibitor (α1PI) varies in the human population and contributes to the risk for GPA manifestations either at onset, during relapses, or during systemic progression. PR3 is initially produced with a signal peptide sequence (a.a. 1-25), an N-terminal and a C-terminal propeptide sequence (a.a. 26-27 and 249-256, respectively), the removal of which yields the mature enzyme (a.a. 28-248).

Clone MCPR3-3 competed against PR3 activity-neutralizing autoantibodies (neutralizing ANCAs) for PR3 binding. The ratios of OD readings obtained with PR3-complexed clone MCPR3-2 (Cat. No. MABT340) to the corresponding ODs obtained with PR3-complexed clone MCPR3-3 in ELISA-based serum ANCA measurements correlate well with the neutralizing activity of ANCAs in patients serum samples (Silva, F., et al. (2010). J. Autoimmun. 35(4) 299-308).

Epitope: On the opposite side (the back side) of the substrates- and α1-PI-binding site (the front side) (Silva, F., et al. (2010). J. Autoimmun. 35(4) 299-308).

Purified human neutrophil PR3.

Anti-Proteinase 3 Antibody, clone MCPR3-3 is an antibody against PR3 for use in ELISA, Flow Cytometry, and Western Blotting.

Research Category
Inflammation & Immunology

Research Sub Category
Inflammation & Autoimmune Mechanisms

Western Blotting Analysis: 4 µg/mL from a representative lot detected 2-5 µg of purified human neutrophil PR3, as well as 1 µg of both pro- and mature forms of recombinant human PR3 under non-reducing condition (Courtesy of Amber Hummel, Mayo Clinic, Rochester, MN).
Western Blotting Analysis: 2-8 µg/mL from a representative lot detected 5 µg of purified human neutrophil PR3 under non-reducing condition, while greatly reduced reactivity was seen upon sample reduction (Courtesy of Amber Hummel, Mayo Clinic, Rochester, MN).
Flow Cytometry Analysis: A representative lot bound immobilized recombinant human PR3 via a distinct epitope than those recognized by clone MCPR3-2 and MCPR3-7 (Cat. No. MABT340 and MABT403, respectively) as determined by FACS analysis of bead-based competition binding assay (Silva, F., et al. (2010). J. Autoimmun. 35(4) 299-308).
Flow Cytometry Analysis: A representative lot bound recombinant human PR3-, but not murine PR3-, coated Talon-beads as determined by FACS analysis. (Silva, F., et al. (2010). J. Autoimmun. 35(4) 299-308).
ELISA Analysis: A representative lot was immobilized on well surface and employed to capture recombinant human PR3, followed by affinity pull-down of PR3 autoantibodies (anti-neutrophil cytoplasmic antibodies or ANCA) from patients serum samples by the captured PR3 and the subsequent detection of the bound ANCA by alkaline phosphatase-conjugated goat anti-human IgG. (Silva, F., et al. (2010). J. Autoimmun. 35(4) 299-308; Sun, J., et al. (1998). J. Immunol. Methods. 211(1-2):111-123).

Identity Confirmation by Isotyping Test.

Isotyping Analysis: The identity of this monoclonal antibody is confirmed by isotyping test to be IgG1κ.

~29 kDa observed. 27.81 kDa (Pro-PR3; a.a. 1-256), 25.14 kDa (N-terminal signal and propeptide removed; a.a. 28-256), 24.25 kDa (Mature; a.a. 28-248) calculated.

Format: Purified

Protein G Purified

Purified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Concentration: Please refer to lot specific datasheet.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.