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Products:Immunology >> Innate Immunity >> Complement >> Classical Pathway
Overview
Product name
Anti-C4d antibody
See all C4d products (7) ...
Description
Rabbit polyclonal to C4d
Tested applications
IHC-P, WB, ICC/IFmore details
Cross reactivity
Reacts with
Human
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 950 - 1050 of Human C4d.
(Peptide available as ab88146.)
Positive control
This antibody gave a positive signal in Human Plasma Total Protein Lysate.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Immunology >> Innate Immunity >> Complement >> Classical Pathway
Applications
Show applications key
Our Abpromise guarantee covers the use of ab87424 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-P: Use a concentration of 5 µg/mlPerform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB: Use a concentration of 1 µg/mlDetects a band of approximately 85 kDa (predicted molecular weight: 193 kDa).
ICC/IF: Use a concentration of 5 µg/ml
Target
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.
Involvement in disease
Defects in C4A are the cause of complement component 4A deficiency (C4AD) [MIM:120810]. A rare defect of the complement classical pathway associated with the development of autoimmune disorders, mainly systemic lupus with or without associated glomerulonephritis.
Sequence similarities
Contains 1 anaphylatoxin-like domain.
Contains 1 NTR domain.
Post-translational
modifications
Prior 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.
N- and O-glycosylated. O-glycosylated with a core 1 or possibly core 8 glycan.
Cellular localization
Secreted.
Target information above from: UniProt accessionP0C0L4
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Acidic complement C4 antibody
- C3 and PZP-like alpha-2-macroglobulin domain-containing protein 2 antibody
- C4 antibody
see all
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
Anti-C4d antibody images:
Western blot - C4d antibody (ab87424)
Anti-C4d antibody (ab87424) at 1 µg/ml + Human Plasma Total Protein Lysate (ab83997) at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution (Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) )
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 193 kDa
Observed band size : 85,193 kDa (why is the actual band size different from the predicted?)
Additional bands at : 110 kDa,130 kDa,30 kDa,48 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 30 seconds
C4d circulates in blood as a disulfide-linked trimer of an alpha, beta and gamma chain. Our immunogen sequence is within the alpha chain. The band at 85kDa could correspond to the alpha chain.
Immunocytochemistry/ Immunofluorescence - C4d antibody (ab87424)
ICC/IF image of ab87424 stained HepG2 cells. The cells were 100% Methanol fixed (5 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 (ab87424, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 100% Methanol fixed (5 min) HeLa, and MCF-7 cells at 5µg/ml.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - C4d antibody (ab87424)
IHC image of C4d staining in human liver carcinoma 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 ab87424, 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.
References for Anti-C4d antibody (ab87424)
ab87424 has not yet been referenced specifically in any publications.
Publishing research using ab87424? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"