Description
The monoclonal antibody S5/1 recognizes the human receptor for C5a (CD88). The C5a receptor is a seven transmembrane GTP-binding-protein-coupled receptor of 45kDa which binds with high affinity to human anaphylatoxin C5a. Upon C5a binding G-protein-dependent cellular responses are initiated such as increased intracellular Ca2+, granule fusion with the cell membrane, enzyme release and oxidative burst suggesting that C5aR is an important mediator of inflammation. The C5a receptor is expressed on a broad range of cell types, including neutrophils, monocytes/macrophages, dendritic cells, astrocytes and microglia.
Interestingly, C5a receptor signalling has been associated with several inflammatory diseases including rheumatoid arthritis.
Clone S5/1 was raised against a synthetic peptide comprising the N-terminal domain of the C5aR (Met1-Asn31). The antibody has been shown to inhibit the binding of C5a to its receptor.
Cross Reactivity
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Cross reactivity
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Cross reactant
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Reactivity
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Rabbit
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Yes9
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Immunogen
Synthetic peptide comprising the N-terminal domain of the C5aR (Met1-Asn31)
Formulation
1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin.
Species
Mouse IgG2a
Application
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F
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FC 1,2,4,7
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FS1,2
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IA
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IF
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IP
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P3,6
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W5
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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
FC: Antibody S5/1 stains the N-terminal domain of CD88. 1x106 leukocytes were incubated with 2µg of antibody (Ref. 1).
FS: Antibody S5/1 inhibits C5a binding to its receptor at a concentration of 3.3µM (Ref. 2)
IHC-P: Tissue sections were fixed in 2% paraformaldehyde and pretreated with 5% normal rabbit serum before applying antibody S5/1 (Ref. 3).
W: extracts from smooth muscle cells were separated by SDS-PAGE, the observed band size with S5/1 antibody is 42kDa (Ref. 5).
Use
For immunohistology, 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 1:10. For inhibition of binding of C5a to its receptor dilutions have to be made according to the amounts of C5a to be inhibited.
Aliases
Complement component 5a receptor 1, CD88, C5aR, C5a anaphylatoxin receptor.
Positive control
Whole blood granulocytes
Negative control
Pulmonary or hepatic epithelial cells
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for one year.
References
1. Opperman, M et al; Probing the human receptor for C5a anaphylatoxin with side-directed antibodies. J Immunol 1993, 151: 3785
2. Opperman, M et al; Plasma clearance of the human C5a anaphylatoxin by binding to leucocyte C5a receptors. Immunol. 1994, 82: 516
3. Werfel, T et al; C5a receptors are detectable on mast cells in normal human skin and in psoriatic plaques but not in weal and flare reactions or in urticaria pigmentosa by immunohistochemistry. Arch Dermatol Res 1997, 289: 83
4. Zwirner, J et al; Evaluation of C3a receptor expression on human leucocytes by the use of novel monoclonal antibodies. Immunology 1999, 97: 166
5. Fortin, J et al; Contractile effect of anaphylatoxin C5a and of a mimetic peptide on the human umbilical artery: further evidence for leukocyte-dependent vasomotion. J Cardiovasc Pharmacol. 2002, 40:815
6. Gueler, F et al; Complement 5a receptor inhibition improves renal allograft survival. J Am Soc Nephrol 2008, 19: 2302
7. Strey, CW et al;. Complement and neutrophil function changes after liver resection in humans. World J Surg 2009, 33: 2635
8. Werfel, T et al: CD88 antibodies specifically bind to C5aR on dermal CD117+ and CD14+ cells and react with a desmosomal antigen in human skin. J Immunol 1996, 157: 1729.
9. Petitclerc, E et al: Pathologic leukocyte infiltration of the rabbit aorta confers a vasomotor effect to chemotactic peptides through cyclooxygenase-derived metabolites. J Immunol 1996, 156: 3426
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 with the use of or derivation of this product.
Also available
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HM2076
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C5/C5a, Human, mAb 561
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HM2077
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C5/C5a, Human, mAb 557
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HM2078
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C5a/C5a des-Arg, Human, mAb 2942
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HM2080
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C5/C5b, Human, mAb 568
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HM2095
|
C5aR, Human, mAb W17/1
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References
1. Opperman, M et al; Probing the human receptor for C5a anaphylatoxin with side-directed antibodies. J Immunol 1993, 151: 3785
2. Opperman, M et al; Plasma clearance of the human C5a anaphylatoxin by binding to leucocyte C5a receptors. Immunol. 1994, 82: 516
3. Werfel, T et al; C5a receptors are detectable on mast cells in normal human skin and in psoriatic plaques but not in weal and flare reactions or in urticaria pigmentosa by immunohistochemistry. Arch Dermatol Res 1997, 289: 83
4. Zwirner, J et al; Evaluation of C3a receptor expression on human leucocytes by the use of novel monoclonal antibodies. Immunology 1999, 97: 166
5. Fortin, J et al; Contractile effect of anaphylatoxin C5a and of a mimetic peptide on the human umbilical artery: further evidence for leukocyte-dependent vasomotion. J Cardiovasc Pharmacol. 2002, 40:815
6. Gueler, F et al; Complement 5a receptor inhibition improves renal allograft survival. J Am Soc Nephrol 2008, 19: 2302
7. Strey, CW et al;. Complement and neutrophil function changes after liver resection in humans. World J Surg 2009, 33: 2635
8. Werfel, T et al: CD88 antibodies specifically bind to C5aR on dermal CD117+ and CD14+ cells and react with a desmosomal antigen in human skin. J Immunol 1996, 157: 1729.
9. Petitclerc, E et al: Pathologic leukocyte infiltration of the rabbit aorta confers a vasomotor effect to chemotactic peptides through cyclooxygenase-derived metabolites. J Immunol 1996, 156: 3426