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Characterization of platelet‐specific alloantigens by immimoblotting: localization of Zw and Bak antigens
Author(s) -
Schoot C. Ellen,
Wester Mieke,
Borne Albert E. G. Kr.,
Huisman Han G.
Publication year - 1986
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1986.tb02233.x
Subject(s) - glycoprotein , microbiology and biotechnology , antibody , platelet , chemistry , platelet membrane glycoprotein , trypsin , antigen , glycoprotein ib , biochemistry , biology , enzyme , immunology
S ummary . The glycoprotein localization of the platelet‐specific antigens Zw a , Zw b and Bak a and their presence on tryptic fragments of glycoproteins was studied by immunoblotting. Human platelets were solubilized and pre‐cleared from platelet‐associated IgG. The glycoproteins were separated on SDS polyacrylamide gels, transferred to nitrocellulose and incubated with platelet antibodies, followed by 125 I‐radiolabelled anti‐human Ig antibodies. Glycoprotein Ilb/IIIa were isolated from platelet lysates by immuno‐affinity chromato‐graphy. These proteins were subjected to trypsin digestion, and then used for the immunoblot procedure with platelet antibodies. A glycoprotein specifically reacting with either anti‐Zw a or anti‐Zw b was found, with an apparent molecular weight of 88 kDa. This protein co‐migrated, and was probably identical with, glycoprotein IIIa. After trypsin digestion the smallest fragment, reactive with IgG anti‐Zw a or IgM anti‐Zw b , had a molecular weight of approximately 23 kDa. IgG anti‐Bak a and anti‐Lek a antibodies reacted with a protein of 130 kDa from platelets of Bak(a+) donors. This protein was identified as glycoprotein IIb.