Measurement of High Affinity Antibodies on Antigen-immunoblots

Overview

Journal J Immunol Methods

Publisher Elsevier

Specialties Allergy & Immunology
Pathology

Date 2006 Feb 7

PMID 16458918

Citations 7

Authors

Miles D Chapman

Geoffrey Keir

Axel Petzold

Edward J Thompson

Affiliations

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Abstract

We describe a semi-quantitative method for measuring the relative affinity of antigen-specific oligoclonal IgG bands separated by isoelectric focusing followed by blotting onto antigen-coated membrane and incubation with sodium thiocyanate. When the developed blot is digitised in greyscale, densitograms can be made and peak areas calculated using ImageJ freeware. By expressing peak area as a percentage of the total area under the curve we have shown that there is a statistically significant rise in percentage of peak area for a given band which persists with increasing molarities of sodium thiocyanate.

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