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MABT340 Anti-Proteinase 3/PR3 Antibody, clone MCPR3-2

Anti-Proteinase 3/PR3 Antibody, clone MCPR3-2 is an antibody against PR3 for use in Affinity Binding Assay, ELISA, Flow Cytometry, and Western Blotting.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Research Category: Cell Structure Research Sub-Category: Inflammation & Autoimmune Mechanisms Gene Symbol: PRTN3, CANCA, MBN, NP4 UniProt Number: P24158
MABT340
100 μg  
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Key Spec Table

Species ReactivityKey ApplicationsHostFormatAntibody Type
HWB, ABA, ELISA, FCMPurifiedMonoclonal Antibody
Description
Catalogue NumberMABT340
DescriptionAnti-Proteinase 3/PR3 Antibody, clone MCPR3-2
Alternate Names
  • Myeloblastin
  • AGP7
  • C-ANCA antigen
  • Leukocyte proteinase 3
  • Neutrophil proteinase 4
  • NP-4
  • P29
  • PR-3
  • PR3
  • Wegener autoantigen
Background InformationMyeloblastin (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).
References
Product Information
FormatPurified
PresentationPurified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Quality LevelMQ100
Applications
ApplicationAnti-Proteinase 3/PR3 Antibody, clone MCPR3-2 is an antibody against PR3 for use in Affinity Binding Assay, ELISA, Flow Cytometry, and Western Blotting.
Key Applications
  • Western Blotting
  • Affinity Binding Assay
  • ELISA
  • Flow Cytometry
Application NotesWestern Blotting Analysis: 2 µg/mL from a representative lot detected 5 µg of purified human neutrophil PR3 under both non-reducing and reducing conditions (Courtesy of Amber Hummel, Mayo Clinic, Rochester, MN).
Flow Cytometry Analysis: A representative lot detected a large population of human peripheral blood neutrophils with surface PR3. The entire population became positively stained after TNFa-stimulation (Hinkofer, L.C., et al. (2013). J. Biol. Chem. 288(37):26635-26648).
Flow Cytometry Analysis: A representative lot bound immobilized recombinant human PR3 via a distinct epitope than those recognized by clone MCPR3-3 and MCPR3-7 (Cat. No. MABF973 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: Representative lots were immobilized on well surface and employed to capture purified human polymorphonuclear cell PR3 or recombinant human PR3 S176A mutant, 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).
Western Blotting Analysis: A representative lot detected an exogenously expressed S176A mutant PR3 in lysates from transfected HMC-1 human mast cells as well as purified PR3 from human polymorphonuclear cells (Sun, J., et al. (1998). J. Immunol. Methods. 211(1-2):111-123).
Affinity Binding Assay: A representative lot captured a recombinant human PR3 construct proP-PR3ctp that adopts a pro-PR3 conformation and a recombinant construct ΔPR3ctp-S195A that adopts a mature PR3 conformation with similar affinity (Hinkofer, L.C., et al. (2013). J. Biol. Chem. 288(37):26635-26648).
Biological Information
ImmunogenGranule extract from HMC-1/PR3 mast cells expressing transfected human neutrophil PR3 (Sun, J., et al. (1998). J. Immunol. Methods. 211(1-2):111-123).
Epitope“North-East” of the substrate-binding pocket and adjacent to clone MCPR3-3 epitope (Silva, F., et al. (2010). J. Autoimmun. 35(4) 299-308).
CloneMCPR3-2
ConcentrationPlease refer to lot specific datasheet.
HostMouse
SpecificityClone MCPR3-2 detected wild-type PR3 and the catalytically inactive S176A PR3 mutant, but not neutrophil elastase or cathepsin G. Clone MCPR3-2 did not interfere with PR3 enzymatic activity or its interaction with α1-protease inhibitor/α1-PI (Sun, J., et al. (1998). J. Immunol. Methods. 211(1-2):111-123). The ratios of OD readings obtained with PR3-complexed clone MCPR3-2 to the corresponding ODs obtained with PR3-complexed clone MCPR3-3 (Cat. No. MABF973) in ELISA-based serum PR3 autoantibodies (ANCAs) measurements correlate well with the neutralizing activity of ANCAs in patients serum samples (Silva, F., et al. (2010). J. Autoimmun. 35(4) 299-308).
IsotypeIgG1κ
Species Reactivity
  • Human
Species Reactivity NoteHuman. Not mouse.
Antibody TypeMonoclonal Antibody
Entrez Gene Number
Gene Symbol
  • PRTN3
  • CANCA
  • MBN
  • NP4
Purification MethodProtein G Purified
UniProt Number
Molecular Weight~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.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality AssuranceIdentity Confirmation by Isotyping Test.

Isotyping Analysis: The identity of this monoclonal antibody is confirmed by isotyping test to be IgG1κ.
Usage Statement
  • 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.
Storage and Shipping Information
Storage ConditionsStable for 1 year at 2-8°C from date of receipt.
Packaging Information
Material Size100 μg
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
MABT340 04055977333596

Documentation

Anti-Proteinase 3/PR3 Antibody, clone MCPR3-2 SDS

Title

Safety Data Sheet (SDS) 

Anti-Proteinase 3/PR3 Antibody, clone MCPR3-2 Certificates of Analysis

TitleLot Number
Anti-Proteinase 3/PR3, -Q2667550 Q2667550
Anti-Proteinase 3/PR3, clone MCPR3-2 - 3208790 3208790
Anti-Proteinase 3/PR3, clone MCPR3-2 - 3308164 3308164
Anti-Proteinase 3/PR3, clone MCPR3-2 - 3328861 3328861
Anti-Proteinase 3/PR3, clone MCPR3-2 - 3739686 3739686
Anti-Proteinase 3/PR3, clone MCPR3-2 - 4083429 4083429
Anti-Proteinase 3/PR3, clone MCPR3-2 - 4149931 4149931
Anti-Proteinase 3/PR3, clone MCPR3-2 - 4194242 4194242
Anti-Proteinase 3/PR3, clone MCPR3-2 - 4208806 4208806
Anti-Proteinase 3/PR3, clone MCPR3-2 Monoclonal Antibody 3109185
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