For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu99 of mouse Crry protein. Antibodies are purified by peptide affinity chromatography.
Crry, also known as complement component receptor 1-like protein (Cr1l), is a murine membrane-bound complement regulatory protein ubiquitously expressed in mouse tissues (1,2). Crry is a functional analog of the human complement decay-accelerating factor (DAF/CD55) and membrane cofactor protein (MCP/CD46) (3). In mice, Crry acts as a cofactor for complement factor I, a serine protease which protects autologous cells against complement-mediated injury by cleaving C3b and C4b deposited on host tissues (4,5). Crry also acts as a decay-accelerating factor, preventing the formation of C4b2a and C3bBb, which are amplification convertases of the complement cascade (3,6). Crry plays a crucial role in early embryonic development by maintaining fetomaternal tolerance (7). On CD4+ T cells, Crry acts as a costimulatory factor which favors IL-4 rather than IFN-gamma secretion, suggesting Th2 polarization (8). Cancer cells and cells of the tumor microenvironment have been shown to overexpress Crry, suppressing tumor-specific T cell responses and complement-mediated anti-tumor immunity (9,10). Multiple isoforms have been identified computationally.
UniProt ID: Q64735
Entrez-Gene Id: 12946
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