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Literature DB >> 22585492

Native flatbed isoelectric focusing for determining antibody clonotype distribution.

Abstract

A sensitive method to analyze specific antibody clonotype changes in a patient with systemic lupus erythematosus, who developed autoantibodies to the Ro 60 autoantigen under observation, is described in this chapter. Patient sera collected over several years were separated by flatbed isoelectric focusing (IEF). Following electrofocusing, the gel is now ready to be analyzed by affinity immunoblotting utilizing Ro 60-coated nitrocellulose membrane to determine oligoclonality of the anti-Ro 60-containing sera.

Entities: Chemical Disease Gene Species

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Year: 2012 PMID： 22585492 PMCID： PMC7310501 DOI： 10.1007/978-1-61779-821-4_21

Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745

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References

12 in total

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Native flatbed isoelectric focusing for determining antibody clonotype distribution.

1. An improved western blotting technique effectively reduces background.

2. Antibody isoelectric spectra visualized by antigen-coated erythrocytes.

3. Isoelectric focusing in polyacrylamide gel and its application to immunoglobulins.

4. Specific characterization of isoelectrofocused immunoglobulins in polyacrylamide gel by reaction with 125 I-labeled protein antigens or antibodies.

5. Affinity immunoblotting. High resolution isoelectric focusing analysis of antibody clonotype distribution.

6. A simple general method of exactly comparing different binding activities and antigenic properties of antibodies by obtaining duplicate copies from a single isoelectric focusing gel.

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