Raising monospecific antibodies by use of protein components prestained with Remazol Brilliant Blue and separated by disc electrophoresis on polyacrylamide gel.

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.

journal_name

Clin Chem

journal_title

Clinical chemistry

authors

Saoji AM,Jad CY,Yemul VL,Khare PM,Kelkar SS

subject

Has Abstract

pub_date

1984-07-01 00:00:00

pages

1252-4

issue

7

eissn

0009-9147

issn

1530-8561

journal_volume

30

pub_type

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