Affinity purification of polyclonal antibodies from antigen immobilized in situ in sodium dodecyl sulfate-polyacrylamide gels

Analytical Biochemistry
P J MadaraI H Mather

Abstract

A procedure for the preparation of affinity-purified antibody is described. Protein mixtures are separated by electrophoresis in sodium dodecyl sulfate (SDS)--polyacrylamide gels. Individual bands of protein are cut from the gel and fixed in situ with glutaraldehyde. The gel pieces are then homogenized and washed extensively with buffered solutions and chaotropic agents. The washed gels can then be used as immunoadsorbents to purify antibodies from crude antisera. This method should be especially useful for the preparation of small amounts of antibody to proteins that are difficult to purify by conventional means, that are available only in limited quantity, or that cannot be blotted to immunoadsorbents such as nitrocellulose or diazotized paper.

References

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