Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4287

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Labeling Antibodies with Iodine

Ed Harlow and David Lane

This protocol was adapted from "Handling Antibodies," Chapter 4, in Using Antibodies by Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA, 1999.

The first 100 words of the full text of this article appear below.


INTRODUCTION

This protocol describes a simple chemical oxidation method for labeling antibodies with iodine. Iodide-125 (supplied as NaI) is oxidized to form iodine-125 (I2), which attacks tyrosyl and histidyl side chains. The iodinated antibodies are easily detected and quantitated using gamma counters or film. They are used primarily in immunoassays, but other techniques can be adapted conveniently to the iodine detection method. Before beginning this procedure, make sure the area in which the iodination will be done is completely prepared and that proper safety precautions have been taken.


MATERIALS

Reagents

recipe caution Antibody (0.2-1 mg/ml in 0.5 M Sodium phosphate, pH 7.5)

caution Carrier-free Na125. . . [Full Text of this Article]

Equipment


METHOD


TROUBLESHOOTING


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