Anti-C3/C3b antibody [EPR19394] ab200999 is a rabbit monoclonal antibody that is used in C3/C3b western blotting and IHC. Suitable for human, mouse and rat samples.
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR19394 is cited in over 90 publications
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
WB | IHC-P | |
---|---|---|
Human | Tested | Tested |
Mouse | Tested | Tested |
Rat | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/2000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/2000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/2000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/2000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/2000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/2000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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C3 plays a central role in the activation of the complement system. Its processing by C3 convertase is the central reaction in both classical and alternative complement pathways. After activation C3b can bind covalently, via its reactive thioester, to cell surface carbohydrates or immune aggregates.Derived from proteolytic degradation of complement C3, C3a anaphylatoxin is a mediator of local inflammatory process. In chronic inflammation, acts as a chemoattractant for neutrophils (By similarity). It induces the contraction of smooth muscle, increases vascular permeability and causes histamine release from mast cells and basophilic leukocytes.C3-beta-cActs as a chemoattractant for neutrophils in chronic inflammation.Acylation stimulating proteinAdipogenic hormone that stimulates triglyceride (TG) synthesis and glucose transport in adipocytes, regulating fat storage and playing a role in postprandial TG clearance. Appears to stimulate TG synthesis via activation of the PLC, MAPK and AKT signaling pathways. Ligand for C5AR2. Promotes the phosphorylation, ARRB2-mediated internalization and recycling of C5AR2 (PubMed:10432298, PubMed:15833747, PubMed:16333141, PubMed:19615750, PubMed:2909530, PubMed:8376604, PubMed:9059512).
CPAMD1, CPAMD1, C3, Complement C3, C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1
Anti-C3/C3b antibody [EPR19394] ab200999 is a rabbit monoclonal antibody that is used in C3/C3b western blotting and IHC. Suitable for human, mouse and rat samples.
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR19394 is cited in over 90 publications
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR19394
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
This supplementary information is collated from multiple sources and compiled automatically.
Complement component 3 (C3) commonly known as C3 complement is a central protein in the complement system which plays a significant role in immune response. C3b a fragment of C3 is produced when C3 undergoes cleavage. C3 is a large protein with a mass of approximately 185 kDa. The liver primarily secretes C3 into the bloodstream. It circulates in the plasma and is found in high concentration making it one of the most abundant components of the complement system.
Complement component C3 forms part of the innate immune system by promoting opsonization which enhances phagocytosis of pathogens. C3b binds to pathogens' surfaces facilitating their recognition by phagocytes. C3 as part of a complex with C3 convertase also has a role in amplifying the activation of the complement cascade. The proteolytic cleavage of C3 into C3b and C3a leads to the activation of other components forming the membrane attack complex and orchestrating inflammation.
The complement component C3 functions within both the classical and alternative complement pathways. It acts as a convergence point where the complement activation pathways meet. C3 is activated into C3b and C3a which are key to amplifying the cascade. Furthermore C3 interacts with proteins such as factor B and factor D in the alternative pathway and C4 and C2 in the classical pathway facilitating the formation of C3 convertase necessary for pathway progression.
Complement C3 shows associations with immune-related and inflammatory diseases. Deficiencies or malfunctions of complement C3 can lead to increased susceptibility to infections due to impaired opsonization and clearance of pathogens. Additionally overactivation of the complement system involving C3 can contribute to autoimmune disorders such as systemic lupus erythematosus. Other proteins linked to these diseases include C4 in lupus and factor H in age-related macular degeneration which controls complement pathway activation.
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We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Full details and terms and conditions can be found here:
Terms & Conditions.
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-C3 antibody [EPR19394] (ab200999) at 1/2000 dilution
Lane 1: Human fetal liver lysate at 10 µg
Lane 2: Human fetal heart lysate at 10 µg
Lane 3: Human fetal kidney lysate at 10 µg
All lanes: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 187 kDa
Observed band size: 115 kDa, 187 kDa, 68 kDa
Exposure time: 30s
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling C3 with ab200999 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Cytoplasm staining on hepatocytes of mouse liver is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1,2,3,5 and 6: 3 minutes; Lane 4,7 and 8: 10 seconds.
All lanes: Western blot - Anti-C3 antibody [EPR19394] (ab200999) at 1/2000 dilution
Lane 1: Mouse brain lysate at 10 µg
Lane 2: Mouse kidney lysate at 10 µg
Lane 3: Mouse spleen lysate at 10 µg
Lane 4: Mouse heart lysate at 10 µg
Lane 5: Rat brain lysate at 10 µg
Lane 6: Rat spleen lysate at 10 µg
Lane 7: Rat heart lysate at 10 µg
Lane 8: Rat kidney lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 187 kDa
Observed band size: 115 kDa, 187 kDa, 43 kDa, 68 kDa
Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling C3 with ab200999 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
The plasma in blood vessel of mouse cardiac muscle was positive staining. The staining pattern is similar to what has been observed in the literature (PMID: 23104558).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling C3 with ab200999 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
The plasma in blood vessels of mouse cerebral cortex was positive staining.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1-5: 8 seconds; Lane 6: 3 minutes.
All lanes: Western blot - Anti-C3 antibody [EPR19394] (ab200999) at 1/2000 dilution
Lane 1: Human serum at 20 µg
Lane 2: Human plasma at 20 µg
Lane 3: HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 4: Mouse plasma at 20 µg
Lane 5: Rat plasma at 20 µg
Lane 6: Human milk at 20 µg
All lanes: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 187 kDa
Observed band size: 115 kDa, 187 kDa, 43 kDa, 68 kDa
Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling C3 with ab200999 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Cytoplasm staining on hepatocytes of rat liver is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling C3 with ab200999 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
The plasma in blood vessels of rat kidney was positive staining. The staining pattern is similar to what has been observed in the literature (PMID: 23104558).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling C3 with ab200999 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
The plasma in blood vessels of rat lung was positive staining. The staining pattern is similar to what has been observed in the literature (PMID: 23104558).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of formalin fixed paraffin embedded human liver labelling C3/C3b with ab200999 at a concentration of 0.01 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.
Anti-C3/C3b antibody [EPR19394] ab200999 was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).
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