Anti-C4d antibody (ab87424)
Key features and details
- Rabbit polyclonal to C4d
- Suitable for: IHC-P, WB, ICC/IF
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-C4d antibody
See all C4d primary antibodies -
Description
Rabbit polyclonal to C4d -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide corresponding to Human C4d aa 950-1050 conjugated to keyhole limpet haemocyanin.
(Peptide available asab88146) -
Positive control
- This antibody gave a positive signal in Human Plasma Total Protein Lysate.
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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 ab87424 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 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. 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. -
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
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. -
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: 120810 Human
- Omim: 120820 Human
- SwissProt: P0C0L4 Human
- SwissProt: P0C0L5 Human
see all -
Alternative names
- acidic C4 antibody
- Acidic complement C4 antibody
- basic C4 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-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.
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Western blot - Anti-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: 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. -
Immunocytochemistry/ Immunofluorescence - Anti-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.
Protocols
Datasheets and documents
References (0)
ab87424 has not yet been referenced specifically in any publications.
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-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.
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Western blot - Anti-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: 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. -
Immunocytochemistry/ Immunofluorescence - Anti-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.
