Protocol

Immunoaffinity Purification: Eluting Antigens from Immunoaffinity Columns

This protocol was adapted from "Immunoaffinity Purification," Chapter 9, in Using Antibodies by Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1999.

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INTRODUCTION

This elution procedure relies on disrupting the bonds between the antibody and the antigen that were created in an earlier procedure (see Immunoaffinity Purification: Binding of Antigen to Antibody-Bead Matrix in a Column or Immunoaffinity Purification: Binding of Antigen to Antibody-Bead Matrix in a Suspension). The conditions may be harsh, denaturing the antibody and the antigen, or mild, leaving both the antigen and antibody in active states. If affinity-purified polyclonal antibodies are being used, the conditions for elution of the antigen will be identical to those used to purify the antibodies. Generally, the optimal . . . [Full Text of this Article]

MATERIALS

Reagents

Equipment

METHOD

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