Nq. Hailat et Sm. Hanash, A TECHNIQUE FOR IDENTIFICATION OF CELLULAR PROTEINS BY WESTERN IMMUNOBLOTTING AND IMMUNOAUTORADIOGRAPHY OF 2-D PAGE, Indian Journal of Biochemistry & Biophysics, 32(5), 1995, pp. 240-244
A major effort of our work has been devoted to the identification, usi
ng two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), of l
ymphoid polypeptides that are involved in lymphoid proliferation or di
fferentiation. We have encountered problems during this effort pertain
ing to the exact localization of a polypeptide(s) in the silver staine
d gels that is recognized by western immunoblotting or proteins detect
ed by autoradiography. In this paper, we present a method, using India
ink stained/immuno-staining replica of 2-D gel nitrocellulose membran
e (NCM) or India ink stained coupled with autoradiography in the case
of phosphoproteins, which allows us to exactly localize the polypeptid
e spots detected by these methods in the silver stained 2-D PAGE. This
method is expected to popularize and widen the use of 2-D PAGE techno
logy in the investigation of cellular polypeptides.