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: 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 Val703 of mouse CD177 protein.
Cluster of differentiation 177 antigen (CD177, also known as human neutrophil alloantigen 2a and NB1), is a glycosyl-phosphatidylinositol (GPI)-linked N-glycosylated transmembrane protein expressed primarily on the surface of neutrophils (1,2). Expression of CD177 can be induced by granulocyte-colony-stimulating factor and during pregnancy or bacterial infection (3-5). CD177 mediates adhesion and transendothelial migration of neutrophils via interaction with CD31/PECAM-1 and integrin β2 on endothelial cells (6,7). CD177 also promotes the cell surface expression of proteinase 3 (PR3), which may play a role in enhancing endothelial cell junctional integrity and thus vascular integrity during neutrophil diapedesis (8-10). In association with CD11b/ITGAM, integrin β2, and PR3, CD177 promotes activation of TNF-α primed neutrophils including degranulation and superoxide production (11). CD177 is a polymorphic gene that has been linked to several important clinical diseases, including polycythemia vera, Wegener’s granulomatosis, and immune mediated neonatal neutropenia (12-14).
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