General description
Complement C3 (UniProt P01024; also known as C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1) is encoded by the C3 (also known as CPAMD1) gene (Gene ID 718) in human. C3 is initially translated with an N-terminal 22-amino acid signal peptide sequence, which is then removed to produce the 1641-amino acid mature C3. It plays a central role in the activation of the complement system. Its activation is required for the activation of both classical and alternative pathways of complement (CPC and APC, respectively). C3 is cleaved into C3a and C3b during CPC activation by the C3-convertase C4b2a composed of the activated C4 and C2. In APC, C3 is cleaved by the C3-convertase C3bBb composed C3b and the activated form of factor B (Bb). C3b serves as an opsonizing agent, and can be further cleaved by Factor I into C3c and C3d. iC3b is a proteolytically inactive C3b fragment that still opsonizes target microbes or cells, but cannot further amplify/activate the complement cascade through APC. iC3b can be further cleaved to C3dg, and finally to C3d. Unregulated activation of APC can result in paroxysmal nocturnal hemoglobinuria (PNH) that is characterized by chronic intravascular hemolysis. Clinical C5-neutralizing mAb treatment prevents the formation of cytolytic membrane attack complex (MAC) of complement, but does not block APC activation. Consequently, PNH patients are left with immune-mediated hemolytic anemia and their erythrocytes become opsonized with complement C3. Monoclonal antibodies (mAbs) against C3b/iC3b are useful for monitoring and studying C3b/iC3b deposit on PNH blood cells and mAbs with neutralizing activities are useful tools for studying C3-mediated CPC and APC.
Specificity
This clone blocks the activation of classical pathway of complement (CPC) by binding C3b and iC3b.
Immunogen
Epitope: iC3b
Sepharose 4B beads with surface C3b/C3bi deposits via APC in normal human serum corresponding to the iC3b of human Complement C3b/iC3b.
Application
Neutralizing Analysis: This clone has been shown to prevent alternative pathway of complement (APC) activation-mediated hemolysis of IgM-opsinized paroxysmal nocturnal hemoglobinuria (PNH) erthyrocytes (Lindorfer, M.A., et al. (2010). Blood. 115(11):2283-2291).
Research Category
Inflammation & Immunology
Research Sub Category
Immunoglobulins & Immunology
This mouse monoclonal Anti-Complement C3b/iC3b Antibody, clone 5G9, Cat. No. MABF961 is a neutralizing antibody validated for use in Flow Cytometry, for the detection of C3b.
Quality
Flow Cytometry Analysis: This antibody (200 ug mAb/5 x 10e6 cells/mL) detected C3b/iC3b deposit on human Burkett′s lymphoma Raji B cells opsonized with anti-CD20 mAb Rituximab (RTX) in the presence of 50% normal human serum (NHS).
Target description
187 kDa calculated
Physical form
Format: Purified
Protein G Purified
Purified mouse monoclonal IgG2aκ antibody in buffer containing PBS without preservatives.
Storage and Stability
Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Other Notes
Concentration: Please refer to lot specific datasheet.
Disclaimer
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.