Technical datasheet
Description
The monoclonal antibody R63 reacts with the rat receptor for C5a (CD88). The rat anaphylatoxin C5a, a 77-amino acid glycopolypeptide generated by proteolytic cleavage of C5, is one of the most potent inflammatory chemo attractants and interacts with two C5a receptors, C5aR and C5L2. Most of the C5a effects are mediated through C5aR. C5aR is known to be crucial in the initiation of acute inflammatory responses. C5a receptor is a seven transmembrane GTP-binding-protein-coupled receptor that belongs to the rhodopsin supergene family. The 45 kDa C5a receptor is expressed by leukocytes such as neutrophils, eosinophils, basophils, monocytes, dendritic cells and mast cells. C5aR can be expressed on both immune and non-immune cells. The latter, include vascular endothelial cells, cardiomyocytes and bronchial epithelial cells. Signaling of C5aR involves intracellular calcium mobilization and activation of among others protein kinase B (PKB) and protein phospholipase D (PLD) pathways. High C5aR expression levels have been associated with the pathogenesis of many inflammatory conditions, autoimmune and neurodegenerative diseases like rheumatoid arthritis, multiple sclerosis, atherosclerosis and inflammatory bowel disease. In vivo blockade of C5aR greatly reduces inflammatory injury. Using immunohistology there is no evidence for non-myeloid expression in healthy control rats. However, under inflammatory situations, the C5aR was found to be upregulated in various organs and tissues including the liver.Antibody
R63 is directed against the amino-terminal domain Ex1 of the rat C5aR.
Immunogen
RBH-2H3 cells transfected with rat C5aR gene
Formulation
1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin and 0.02% sodium azide.
Species
Mouse IgG1
Application
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F1,2,5-6
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FC1,2, 3
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FS2
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IA2
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IF1,3,4
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IP
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P
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W5,6
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Yes
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No
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N.D.
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N.D.= Not Determined; F = Frozen sections; FC = Flow Cytometry; FS = Functional Studies; IA = Immuno Assays; IF = Immuno Fluorescence; IP = Immuno Precipitation; P = Paraffin sections; W = Western blot
Application notes
IHC-F: 7µm frozen paraformaldehyde fixed sections were blocked with serum and incubated with mAb at 5 µg/ml
FC: 2x105 cells were incubated with mAb at 3-5µg/ml, all incubations on ice
W: cell homogenate was separated on 10% SDS-PAGE and blotted on nitrocellulose. After blocking blot was incubated with mAb (1/500-1000)
Use
For immunohistochemistry, flow cytometry and Western blotting, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.
Aliases
C5aR; CD88; C5a anaphylatoxin chemotactic receptor; C5a anaphylatoxin receptor; C5a ligand; C5a-R; complement component 5a receptor 1
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least one year. The exact expiry date is indicated on the label.
References
1. Schlaf, G et al; Differential expression of the C5a receptor on the main cell types of rat liver as demonstrated with a novel monoclonal antibody and by C5a anaphylatoxin-induced Ca2+ release. Lab Invest 1999, 79: 1287
2. Rothermel, E et al; Analysis of the tissue distribution of the rat C5a receptor and inhibition of C5a-mediated effects through the use of two MoAbs. Scand J Immunol 2000, 52: 401
3. Schieferdecker, H et al; Induction of functional anaphylatoxin C5a receptors on hepatocytes by in vivo treatment of rats with IL-6. J Immunol 2000, 164: 5453
4. Griffin, R et al; Complement induction in spinal cord microglia results in anaphylatoxin C5a-mediated pain hypersensitivity. J. Neurosc. 2007, 27: 8699
5. Woodruff, T et al; The complement factor C5a contributes to pathology in a rat model of amyoptrophic lateral sclerosis. J Immunol 2008, 181: 8727
6. Crane, J et al; The C5a anaphylatoxin receptor CD88 is expressed in presynaptic terminals of hippocampal mossy fibres. J Neuroinflamation 2009, 6: 34
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result from the use or derivation of this product.
Also available
References
1. Schlaf, G et al; Differential expression of the C5a receptor on the main cell types of rat liver as demonstrated with a novel monoclonal antibody and by C5a anaphylatoxin-induced Ca2+ release. Lab Invest 1999, 79: 1287
2. Rothermel, E et al; Analysis of the tissue distribution of the rat C5a receptor and inhibition of C5a-mediated effects through the use of two MoAbs. Scand J Immunol 2000, 52: 401
3. Schieferdecker, H et al; Induction of functional anaphylatoxin C5a receptors on hepatocytes by in vivo treatment of rats with IL-6. J Immunol 2000, 164: 5453
4. Griffin, R et al; Complement induction in spinal cord microglia results in anaphylatoxin C5a-mediated pain hypersensitivity. J. Neurosc. 2007, 27: 8699
5. Woodruff, T et al; The complement factor C5a contributes to pathology in a rat model of amyoptrophic lateral sclerosis. J Immunol 2008, 181: 8727
6. Crane, J et al; The C5a anaphylatoxin receptor CD88 is expressed in presynaptic terminals of hippocampal mossy fibres. J Neuroinflamation 2009, 6: 34