Semi-dry protein transfer and immunodetection of P-selectin using an antibody to its C-terminal tag.
P-selectin is a multidomain glycoprotein expressed on activated endothelial cells. We previously expressed a recombinant form of P-selectin containing only its N-terminal lectin and EGF domains in CHO-K1 cells and showed that these two domains are sufficient to mediate ligand binding. We have now expressed the same construct in CHO-Lec1 cells that make truncated glycans. The uniform glycosylation in these cells should make it easier to crystallize this protein.
Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, 73104, USA. Padmaja-Mehta@omrf.org
Padmaja
P
Mehta
D000818:Animals; D000906:Antibodies / Q000276:immunology; D015153:Blotting, Western / Q000295:instrumentation / Q000379:methods*; D002460:Cell Line; D019007:P-Selectin / Q000737:chemistry* / Q000235:genetics; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics
D000906:Antibodies; D019007:P-Selectin; D011994:Recombinant Proteins