Complement C3 Antibody (11H9) - BSA Free

Flow (Intracellular): Complement C3 Antibody (11H9) [NB200-540] - An intracellular stain was performed on RAW 246.7 cells with Complement C3 (11H9-3-2) antibody NB200-540 (blue) and a matched isotype control NBP2-31382 (orange). Cells were either treated with 3uM Monensin for 3 hours to block the secretion of Complement C3 (B) or grown in normal media (A). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2 ug/mL for 30 minutes at room temperature, followed by mouse F(ab)2 IgG (H+L) PE-conjugated secondary antibody (F0102B, R&D Systems).

Immunocytochemistry/Immunofluorescence: Complement C3 Antibody (11H9) [NB200-540] - C3 protein fragments deposited on kidney cells of MPL-lpr mouse. Staining with antibody 11H9. Glomerular staining pattern. Fixation in 4% paraformaldehyde in PBS pH 7.4. Vibratome sections of 4 um. Pretreated with 3% hydrogen peroxide for 20 min to quench endogenous peroxidases. Microwaved in antigen unmasking solution for 2-5 minutes as antigen retrieval.

Immunohistochemistry-Paraffin: Complement C3 Antibody (11H9) [NB200-540] - Complement C3 protein in a FFPE tissue section of mouse liver using 1:100 dilution of Complement C3 antibody (clone 11H9) NB200-540. Weak but distinct membrane-cytoplasmic immunopositivity was observed in hepatocytes and few cells developed punctate membrane staining.

Flow Cytometry: Complement C3 Antibody (11H9) [NB200-540] - Left panel: FMO, Middle panel: No primary antibody control, Right panel: sample. Day 6 murine mammary tumors processed and stained for analysis with flow cytometry. The C3b+ population of CD45+ cells is what the gate in each sample is exhibiting. WB image submitted by a verified customer review.

Immunohistochemistry-Paraffin: Complement C3 Antibody (11H9) [NB200-540] - Complement C3 protein in a FFPE tissue section of mouse lymph node using 1:100 dilution of Complement C3 antibody (clone 11H9) NB200-540. This representative photomicrograph shows a membrane-cytoplasmic immunopositivity in non-germinal center cells, and few cells developed an intense staining for this target protein.

• Reactivity: Mu, Ec
• Applications: Flow, Flow-IC, IA, ICC/IF, IHC, IHC-Fr, IHC-P, IP, CyTOF-ready
• Clone: 11H9
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

Complement C3 Antibody (11H9) - BSA Free Summary

• Immunogen: This Complement C3 Antibody (11H9) was developed against C57BL/6 thymocytes saturated with rat anti-Thy-1 monoclonal antibody of IgG2b subclass (RmT1).
• Localization: Secreted
• Specificity: Mouse Complement C3 and its activation products, C3b, iC3b, C3d and C3dg
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Rat
• Gene: C3
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow (Intracellular)
  • Flow Cytometry 1:10-1:1000
  • Immunoassay 0.5 ug/well in PBS
  • Immunocytochemistry/Immunofluorescence 1:10-1:500
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:10-1:500
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Immunoprecipitation 1:10-1:500
• Theoretical MW: 187 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Use in Mouse reported in scientific literature (PMID:34433493). Use in E. coli reported in scientific literature (PMID:32422907).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for Complement C3 Antibody (11H9) - BSA Free

• Acylation Stimulating Protein
• acylation-stimulating protein cleavage product
• AHUS5
• ARMD9
• ASP
• C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1
• C3
• C3a anaphylatoxin
• C3a
• C3adesArg
• C3b
• C3bc
• C3-beta-c
• Complement C3 alpha chain
• Complement C3 beta chain
• complement C3
• Complement C3b alpha' chain
• Complement C3c alpha' chain fragment 1
• Complement C3c alpha' chain fragment 2
• Complement C3d fragment
• Complement C3dg fragment
• Complement C3f fragment
• Complement C3g fragment
• complement component 3
• complement component C3
• complement component C3a
• complement component C3b
• CPAMD1
• EC 3.4.21.43
• epididymis secretory sperm binding protein Li 62p
• HEL-S-62p
• prepro-C3

Background

The complement system, or complement cascade, is a part of the innate immune system that assists in defense against pathogens (1-3). Complement C3, also called C3 or C3 protein, is one of nine complement proteins and is the main component of the complement system which is composed of over 30 soluble and membrane-bound proteins (1,4). The complement cascade consists of three main pathways: the classical, the lectin, and the alternative, all of which converge into a common pathway involving C3 cleavage by C3-convertases (1-6). Human Complement C3 is synthesized as a protein of 1663 amino acids (aa) in length with a theoretical molecular weight of ~185 kDa (5). Complement C3 is the most prevalent human complement protein in the serum, with a concentration of 1.2 mg/mL, and is predominantly produced by hepatocytes in the liver, but is also synthesized by blood cells and epithelial cells (3,5). Furthermore, the structure of C3 is comprised of an alpha-chain (110 kDa) and a beta-chain (75 kDa) linked by a disulfide bond (5). Cleavage of inactive C3 by C3-convertases produces active C3a, which functions as a mediator of inflammation, and C3b, which is an opsonin (1-4). In addition to amplification of complement response, C3 fragments serve multiple additional functions including chemotaxis, phagocytosis, adhesion, and immune modulation (3). Complement C3 serves dual purposes where it is involved in pathogenesis and immunity but, conversely, cellular damage results from unregulated C3 activation (5).

Both elevated levels and reduced levels of Complement C3 has been implicated in diseases pathologies (6). Deficiency in Complement proteins can result in autoimmune disorders including systemic lupus erythematosus, which is more often associated with C1 or C4 deficiency and only rarely with C3 deficiency (6). However, C3 deficiency typically results in increased risk of recurrent bacterial infections and glomerulonephritis, characterized by inflammation of the filtering glomeruli in the kidneys (6). Additionally, elevated levels of C3a and C4a is seen in patients with antiphospholipid antibody syndrome (6). Serum levels of C3 are also higher in rheumatoid arthritis cases (6). The complement system has become a target for drugs and therapeutics aimed at modulating innate immunity (7). For instance, compstatin is a peptide that binds to C3, inhibiting convertase activity and cleavage and can be used to treat diseases associated with uncontrolled C3 activation (7). C3-inhibitors and other complement inhibitors are a promising drug candidate for treatment of many diseases (7).

References

1. Mathern, D. R., & Heeger, P. S. (2015). Molecules Great and Small: The Complement System. Clinical Journal of the American Society of Nephrology: CJASN. https://doi.org/10.2215/CJN.06230614

2. Merle, N. S., Church, S. E., Fremeaux-Bacchi, V., & Roumenina, L. T. (2015). Complement System Part I - Molecular Mechanisms of Activation and Regulation. Frontiers in Immunology. https://doi.org/10.3389/fimmu.2015.00262

3. Ricklin, D., Reis, E. S., Mastellos, D. C., Gros, P., & Lambris, J. D. (2016). Complement component C3 - The "Swiss Army Knife" of innate immunity and host defense. Immunological Reviews. https://doi.org/10.1111/imr.12500

4. Merle, N. S., Noe, R., Halbwachs-Mecarelli, L., Fremeaux-Bacchi, V., & Roumenina, L. T. (2015). Complement System Part II: Role in Immunity. Frontiers in Immunology. https://doi.org/10.3389/fimmu.2015.00257

5. Sahu, A., & Lambris, J. D. (2001). Structure and biology of complement protein C3, a connecting link between innate and acquired immunity. Immunological Reviews. https://doi.org/10.1034/j.1600-065x.2001.1800103.x

6. Vignesh, P., Rawat, A., Sharma, M., & Singh, S. (2017). Complement in autoimmune diseases. Clinica Chimica Acta; International Journal of Clinical Chemistry. https://doi.org/10.1016/j.cca.2016.12.017

7. Mastellos, D. C., Yancopoulou, D., Kokkinos, P., Huber-Lang, M., Hajishengallis, G., Biglarnia, A. R., Lupu, F., Nilsson, B., Risitano, A. M., Ricklin, D., & Lambris, J. D. (2015). Compstatin: a C3-targeted complement inhibitor reaching its prime for bedside intervention. European Journal of Clinical Investigation. https://doi.org/10.1111/eci.12419

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Complement C3 Antibody (NB200-540)(15)

Publications using NB200-540

• Liu T, Yang M, Xia Y Et al. Microarray-based analysis of renal complement components reveals a therapeutic target for lupus nephritis Arthritis research & therapy Aug 25 2021 [PMID: 34433493] (IHC-Fr, Mouse)
• Davidson S, Efremova M, Riedel A et al. Single-Cell RNA Sequencing Reveals a Dynamic Stromal Niche That Supports Tumor Growth Cell Rep May 19 2020 [PMID: 32433953] (ICC/IF, Mouse)
• Holland-Tummillo KM, Shoudy LE, Steiner D et al. Autotransporter-Mediated Display of Complement Receptor Ligands by Gram-Negative Bacteria Increases Antibody Responses and Limits Disease Severity Pathogens May 14 2020 [PMID: 32422907] (IP, E. coli)
• Sun D, Sun P, He S, Shi M Rat IgG mediated circulatory cell depletion in mice requires mononuclear phagocyte system and is facilitated by complement J. Leukoc. Biol. Jan 22 2020 [PMID: 31965640] (Flow, Mouse)
• Wang K Wei H Zhan J et al. GSPE alleviates renal fibrosis by inhibiting the activation of C3/ HMGB1/ TGF- beta 1 pathway Chem. Biol. Interact. Dec 23 2019 [PMID: 31874164] (ICC/IF, Mouse)
• Campbell SC, MuNoz-Ballester C, Chaunsali L, et al. Potassium and glutamate transport is impaired in scar-forming tumor-associated astrocytes Neurochem. Int. Dec 9 2019 [PMID: 31825815] (IHC, Mouse)
• Wu T, Dejanovic B, Gandham VD et al. Complement C3 Is Activated in Human AD Brain and Is Required for Neurodegeneration in Mouse Models of Amyloidosis and Tauopathy Cell Rep Aug 20 2019 [PMID: 31433986] (IHC, Mouse)
• Medler TR, Murugan D, Horton W et al. Complement C5a Fosters Squamous Carcinogenesis and Limits T Cell Response to Chemotherapy. Cancer Cell. Oct 8 2018 [PMID: 30300579] (IHC, Mouse)
• Maehara N, Arai S, Mori M et al. Circulating AIM Prevents Hepatocellular Carcinoma through Complement Activation. Cell Rep. 2014 Oct 09 [PMID: 25284781] (IHC, Mouse)
• Davidson S Defining the pro-tumour impact of the evolving stromal microenvironment Thesis Jan 1 2020
• Griffin P, Sheehan PW, Dimitry JM et al. REV-ERB alpha mediates complement expression and diurnal regulation of microglial synaptic phagocytosis eLife Dec 1 2020 [PMID: 33258449] (IHC, Mouse)
• Chakraborty S Complement factors in murine jejunum mucus and their functional alteration after experimental sepsis Thesis Jan 1 2021 (ELISA, Mouse)
• Lee SH, Meilandt WJ, Xie L et al. Trem2 restrains the enhancement of tau accumulation and neurodegeneration by beta-amyloid pathology Neuron Feb 27 2021 [PMID: 33675684]
• Liu T, Yang M, Xia Y et al. Microarray-Based Analysis of the Complements in Kidney Reveals a Therapeutic Target in Lupus Nephritis Research Square Mar 26 2021
• Yang L, Li N, Yang D et al. CCL2 regulation of MST1-mTOR-STAT1 signaling axis controls BCR signaling and B-cell differentiation Cell death and differentiation Apr 20 2021 [PMID: 33879857]

Reviews for Complement C3 Antibody (NB200-540) (2)

Average Rating: 4.5

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Mouse.

reviewed by: Francesca Azar Flow Mouse 05/16/2019

Summary

• Application: Flow Cytometry
• Sample Tested: Breast tumor
• Species: Mouse
• Lot: AB042114A-06

Comments

• Comments: Despite some issues with our secondary antibody, this antibody worked well for flow cytometry analysis of mouse tumor tissue.

reviewed by: Verified Customer ICC Mouse 04/01/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: brain and spinal cord
• Species: Mouse

FAQs for Complement C3 Antibody (NB200-540). (Showing 1 - 1 of 1 FAQs).

1. I am trying to establish a method to measure mice C3 levels by nephelometry. I would be most grateful if you could provide me with some help, regarding the choice of the Ab. I totally understand that since such a method has never been tried, I do not expect any guaranties. 
   • We have never performed nephelometry in our lab, and do not have a protocol or advice to provide about this application. However, it seems to me that you should choose an antibody that is capable of recognizing its target in its folded conformation. Therefore, I would suggest trying an antibody that has been validated for ICC or IHC.

Bioinformatics

• Gene Symbol: C3
• Entrez:
  • 12266(Mouse)
• Uniprot:
  • P01027()