Key features and details
- Rabbit polyclonal to C4d
- Suitable for: IHC-P, WB, ICC/IF
- Reacts with: Human
- Isotype: IgG
Product nameAnti-C4d antibody
See all C4d primary antibodies
DescriptionRabbit polyclonal to C4d
Tested applicationsSuitable for: IHC-P, WB, ICC/IFmore details
Species reactivityReacts with: Human
Synthetic peptide corresponding to Human C4d aa 950-1050 conjugated to keyhole limpet haemocyanin.
(Peptide available as
- This antibody gave a positive signal in Human Plasma Total Protein Lysate.
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
Immunizing Peptide (Blocking)
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/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 85 kDa (predicted molecular weight: 193 kDa).|
|ICC/IF||Use a concentration of 5 µg/ml.|
FunctionC4 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 diseaseDefects 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 similaritiesContains 1 anaphylatoxin-like domain.
Contains 1 NTR domain.
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.
N- and O-glycosylated. O-glycosylated with a core 1 or possibly core 8 glycan.
- Information by UniProt
- acidic C4 antibody
- Acidic complement C4 antibody
- basic C4 antibody
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.
Anti-C4d antibody (ab87424) at 1 µg/ml + Human Plasma Total Protein Lysate (ab83997) at 10 µg
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: 193,85 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.
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.
ab87424 has not yet been referenced specifically in any publications.