Complement C3 Antibody (JF10-30)

Novus Biologicals | Catalog # NBP2-66994

Recombinant Monoclonal Antibody.
Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Mouse

Applications

Validated:

Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # JF10-30 expressed in HEK293
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Product Specifications

Immunogen

Synthetic peptide within Human Complement C3 aa 1,210-1,248 / 1,663. (SwissProt: P01024 Human; SwissProt: P01027 Mouse)

Localization

Secreted.

Specificity

Can detect the following chains: Complement C3 alpha chain, Complement C3b alpha, Complement C3dg, and Complement C3d

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

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.

Description

Novus Biologicals Rabbit Complement C3 Antibody (JF10-30) (NBP2-66994) is a recombinant monoclonal antibody validated for use in IHC, WB, Flow and ICC/IF. Anti-Complement C3 Antibody: Cited in 1 publication. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for Complement C3 Antibody (JF10-30)

Immunocytochemistry/ Immunofluorescence: Complement C3 Antibody (JF10-30) [NBP2-66994]

Immunocytochemistry/ Immunofluorescence: Complement C3 Antibody (JF10-30) [NBP2-66994]

Immunocytochemistry/Immunofluorescence: Complement C3 Antibody (JF10-30) [NBP2-66994] - Staining C3 in NIH/3T3 cells (red). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X-100, PBS.
Flow Cytometry: Complement C3 Antibody (JF10-30) [NBP2-66994]

Flow Cytometry: Complement C3 Antibody (JF10-30) [NBP2-66994]

Flow Cytometry: Complement C3 Antibody (JF10-30) [NBP2-66994] - Analysis of HepG2 cells with C3 antibody at 1:50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody
Complement C3 Antibody (JF10-30)

Flow Cytometry: Complement C3 Antibody (JF10-30) [NBP2-66994] -

Flow Cytometry: Complement C3 Antibody (JF10-30) [NBP2-66994] - Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4c for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4 c. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Complement C3 Antibody (JF10-30)

Western Blot: Complement C3 Antibody (JF10-30) [NBP2-66994] -

Western Blot: Complement C3 Antibody (JF10-30) [NBP2-66994] -Western blot analysis of C3 on different lysates with Rabbit anti-C3 antibody at 1/1,000 dilution.Lane 1: Human liver tissue lysateLane 2: HepG2 cell lysateLane 3: HeLa cell lysateLane 4: Mouse liver tissue lysateLane 5: Mouse spleen tissue lysateLane 6: Rat liver tissue lysateLane 7: Rat spleen tissue lysateLysates/proteins at 30 µg/Lane.Predicted band size: 187 kDaObserved band size: 108/37 kDaExposure time: 1 minute;4-20% SDS-PAGE gel.
Complement C3 Antibody (JF10-30)

Immunocytochemistry/Immunofluorescence: Complement C3 Antibody (JF10-30) [NBP2-66994] -

Immunocytochemistry/Immunofluorescence: Complement C3 Antibody (JF10-30) [NBP2-66994] -Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-C3 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4 c. Goat Anti-Mouse IgG H&L (iFluor™ ) was used as the secondary antibody at 1/1,000 dilution.

Applications for Complement C3 Antibody (JF10-30)

Application
Recommended Usage

Flow Cytometry

1:50-1:100

Immunocytochemistry/ Immunofluorescence

1:50-1:100

Immunohistochemistry-Paraffin

Reported in scientific literature (PMID: 31578522)

Western Blot

1:1000

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Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

TBS (pH7.4), 0.05% BSA, 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Complement C3

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

Alternate Names

Acylation Stimulating Protein, acylation-stimulating protein cleavage product, AHUS5, ARMD9, ASP, C3, C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1, C3a, C3a anaphylatoxin, C3adesArg, C3b, C3bc, C3-beta-c, complement C3, Complement C3 alpha chain, Complement C3 beta chain, 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

Gene Symbol

C3

Additional Complement C3 Products

Product Documents for Complement C3 Antibody (JF10-30)

Certificate of Analysis

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Product Specific Notices for Complement C3 Antibody (JF10-30)

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.

Citations for Complement C3 Antibody (JF10-30)

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Complement C3 Antibody (JF10-30)

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  • Q: 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. 

    A: 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.

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