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Catalog # HM1076

C5aR, Mouse, mAb 20/70

120,00561,00

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The monoclonal antibody 20/70 recognizes the mouse receptor for C5a (CD88). The mouse anaphylatoxin complement factor C5a, a split product of C5, is one of the most potent inflammatory chemoattractants and interacts with two C51 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 nonimmune cells. Nonimmune cells expressing C5aR 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 diseases. In vivo blockade of C5aR greatly reduces inflammatory injury. The monoclonal antibody 20/70 has been used in functional studies and is capable to prevent the binding of C5a to its receptor.

Application
Flow cytometry, Functional studies, Immuno fluorescence

Application Notes
FC: 2x10^5 cells were blocked with human and murine IgG on ice for 20’. Cells were stained with 10μg antibody 20/70 for 45’ in PBS/1.5S. For analysis with FACS cells were washed with PBS/1%formaldehyde (Ref.2).
FS: Before migration towards C5a was induced, 2x105 cells were incubated for 30’ with 10μg antibody 20/70 in a transwell migration assay (Ref.2).
IF: Cells were cultured on coverslips, fixed with 4% PFA for 20’, blocked with 3% BSA for 2h and stained with antibody 20/70 for 2h (Ref.6).

Use
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. For functional studies, in vitro dilutions have to be optimized in user’s experimental setting.

Positive Control
Mouse neutrophils

Negative Control
HEK293 cells

Product type
Monoclonal antibodies
Quantity
20 µg, 100 µg
Formulation
0.2 ml (100 μg/ml) 0.2 μm protein G purified antibody solution in PBS, containing 0.1% bovine serum albumin.
Immunogen
RBL-2H3 transfectantsexpressing murine C5aR on the cell surface.
Isotype
Rat IgG2b
Species
Mouse
Alias
C5a anaphylatoxin chemotactic receptor 1, C5a anaphylatoxin chemotactic receptor, C5a-R<br> - Gene name: C5ar1
Storage and stability
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for one year.
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.
Disease
Infectious diseases, Nephrology

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