Remazol Brilliant Blue as a pre-stain for the immediate visualization of human serum proteins on polyacrylamide gel disc electrophoresis.

1983 ◽  
Vol 29 (1) ◽  
pp. 42-44 ◽  
Author(s):  
A M Saoji ◽  
C Y Jad ◽  
S S Kelkar

Abstract We describe the pre-staining of proteins in normal human serum with Remazol Brilliant Blue before separation by disc electrophoresis. Critical to reproducibility are: dye concentration of 0.16 mol/L in a Tris-glycine buffer (pH 8.3), use of equal volumes of serum and dye solution, a tagging period of 2 h at room temperature, and electrophoresis of 0.1 mL of the mixture at 2.5 mA/gel for about 2 h. Advantages include speed, quality of resolution, and low background. This method was compared with Amido Black post-electrophoresis staining in 35 sera. Of these, 16 showed identical results with respect to the number of bands. In the other 19, Amido Black showed more bands in the post-transferrin region and, sometimes, in the post-albumin region. The pre-stained gels showed slower electrophoretic mobilities of the components. Protein bands eluted from pre-stained gels retained immunological reactivity.

1984 ◽  
Vol 30 (7) ◽  
pp. 1252-1254 ◽  
Author(s):  
A M Saoji ◽  
C Y Jad ◽  
V L Yemul ◽  
P M Khare ◽  
S S Kelkar

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.


1971 ◽  
Vol 68 (1_Supplb) ◽  
pp. S105 ◽  
Author(s):  
O. A. Lea ◽  
K. F. Støa

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