Anti-C4b antibody (ab66791)
Key features and details
- Rabbit polyclonal to C4b
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-C4b antibody
See all C4b primary antibodies -
Description
Rabbit polyclonal to C4b -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Chimpanzee, Non human primates
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1100 - 1200 of Human C4b.
Read Abcam's proprietary immunogen policy (Peptide available as ab74629.) -
Positive control
- This antibody gave a positive signal in Human Plasma Total Protein IHC-P: FFPE Human Liver tissue sections.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab66791 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application Abreviews Notes ICC/IF Use a concentration of 10 µg/ml. IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. WB Use a concentration of 1 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 90 , 180 kDa). Target
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Function
C4 plays a central role in the activation of the classical pathway of the complement system. It is processed by activated C1 which removes from the alpha chain the C4a anaphylatoxin. The remaining alpha chain fragment C4b is the major activation product and is an essential subunit of the C3 convertase (C4b2a) and the C5 convertase (C3bC4b2a) enzymes of the classical complement pathway.
Derived from proteolytic degradation of complement C4, C4a anaphylatoxin is a mediator of local inflammatory process. It induces the contraction of smooth muscle, increases vascular permeability and causes histamine release from mast cells and basophilic leukocytes. -
Sequence similarities
Contains 1 anaphylatoxin-like domain.
Contains 1 NTR domain. -
Post-translational
modificationsPrior to secretion, the single-chain precursor is enzymatically cleaved to yield the non-identical chains (alpha, beta and gamma). During activation, the alpha chain is cleaved by C1 into C4a and C4b, and C4b stays linked to the beta and gamma chains. Further degradation of C4b by C1 into the inactive fragments C4c and C4d blocks the generation of C3 convertase. -
Cellular localization
Secreted. - Information by UniProt
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Database links
- Entrez Gene: 100293534 Human
- Entrez Gene: 100507685 Human
- Entrez Gene: 720 Human
- Entrez Gene: 721 Human
- Omim: 120820 Human
- SwissProt: P0C0L4 Human
- SwissProt: P0C0L5 Human
- Unigene: 534847 Human
see all -
Alternative names
- Basic complement C4 antibody
- C2B5 antibody
- C3 and PZP-like alpha-2-macroglobulin domain-containing protein 3 antibody
see all
Images
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Western blot - Anti-C4b antibody (ab66791)All lanes : Anti-C4b antibody (ab66791) at 1 µg/ml
Lane 1 : Human Plasma Total Protein at 2 µg
Lane 2 : Human Plasma Total Protein at 5 µg
Lane 3 : Human Plasma Total Protein at 10 µg
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 90 , 180 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutes
We hypothesize that the 90 kDa band represents the C4b-A polypeptide chain, which is a cleavage product of C4b. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C4b antibody (ab66791)IHC image of ab66791 staining in human normal liver* formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab66791, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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Immunocytochemistry/ Immunofluorescence - Anti-C4b antibody (ab66791)ICC/IF image of ab66791 stained HepG2 cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66791, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Protocols
Datasheets and documents
References (1)
ab66791 has been referenced in 1 publication.
- Heo SH et al. Fabry disease: characterisation of the plasma proteome pre- and post-enzyme replacement therapy. J Med Genet 54:771-780 (2017). PubMed: 28835480
Images
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Western blot - Anti-C4b antibody (ab66791)All lanes : Anti-C4b antibody (ab66791) at 1 µg/ml
Lane 1 : Human Plasma Total Protein at 2 µg
Lane 2 : Human Plasma Total Protein at 5 µg
Lane 3 : Human Plasma Total Protein at 10 µg
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 90 , 180 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutes
We hypothesize that the 90 kDa band represents the C4b-A polypeptide chain, which is a cleavage product of C4b. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C4b antibody (ab66791)
IHC image of ab66791 staining in human normal liver* formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab66791, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
-
Immunocytochemistry/ Immunofluorescence - Anti-C4b antibody (ab66791)ICC/IF image of ab66791 stained HepG2 cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66791, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
