Prestaining proteins

T. S. Pillay tpillay at ucsd.edu
Fri Jun 24 08:06:23 EST 1994


A couple of months ago, I posted a method for prestaining proteins and
some people have written to say they cannot get it to work.  I enclose
the original references (and a few more) which may be of help towards
optimising the conditions.

-Saoji AM; Jad CY; Kelkar SS.
     Remazol Brilliant Blue as a pre-stain for the immediate
visualization of
     human serum proteins on polyacrylamide gel disc electrophoresis.
   Clinical Chemistry, 1983 Jan, 29(1):42-4.
     ABSTRACT available.  (UI:  83077593)

-Deo SS; Fule RP; Saoji AM.
     Drimarene brilliant blue--a better pre-stain in protein separation
and
     visualization.
   Indian Journal of Experimental Biology, 1990 Dec, 28(12):1184-6.
     ABSTRACT available.  (UI:  91285726)

-Saoji AM; Jad CY; Yemul VL; Khare PM; Kelkar SS.
     Raising monospecific antibodies by use of protein components
prestained
     with Remazol Brilliant Blue and separated by disc electrophoresis on
     polyacrylamide gel.
   Clinical Chemistry, 1984 Jul, 30(7):1252-4.
     ABSTRACT available.  (UI:  84234346)


 

1. Saoji AM; Jad CY; Yemul VL; Khare PM; Kelkar SS.
     Raising monospecific antibodies by use of protein components
prestained
     with Remazol Brilliant Blue and separated by disc electrophoresis on
     polyacrylamide gel.
   Clinical Chemistry, 1984 Jul, 30(7):1252-4.
     (UI:  84234346)

Abstract: We have reported (Clin Chem 29: 42-44, 1983) that prestaining
with
    Remazol Brilliant Blue permits direct visualization of serum
components on
    disc electrophoresis, and apparently purifies the proteins well. Here
we
    have cut out the bands corresponding to the prestained albumin and
    transferrin after disc electrophoresis of normal human serum proteins,
    eluted some individual proteins into saline, and assessed their
purity by
    immunoelectrophoresis and two-dimensional crossed
immunoelectrophoresis
    against polyvalent antihuman serum. These two techniques indicated
purity
    of these antigens. We inoculated rabbits with the eluates containing
the
    pure antigens, and tested the resulting antibodies for
monospecificity by
    immunoelectrophoresis, rocket electrophoresis, and single radial
    immunodiffusion. From the results we conclude that the antibodies
raised
    against each component were monospecific, and that this is a simple,
    economical, rapid, and reliable method for obtaining a pure fraction
of
    serum protein for use as an antigen.



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