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.
Mehta
Padmaja
P
Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, 73104, USA. Padmaja-Mehta@omrf.org
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