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Affinity immunoblotting - High resolution isoelectric focusing analysis of antibody clonotype distributionA sensitive and specific method is proposed for the analysis of specific antibody clonotype changes occurring during an immune response and for comparing multiple sera for antibody clonotype similarities. Polyclonal serum antibodies separated by isoelectric focusing (IEF) were analyzed by an affinity immunoblotting method using antigen-coated nitrocellulose membranes. Antibodies present on the surface of the acrylamide gels following IEF bind the antigen on the nitrocellulose when the coated nitrocellulose is laid over the gels. The technique has been used to analyze Ig clonotypes specific for five protein antigens and two carbohydrate antigens. Optimal antigen concentrations for coating the nitrocellulose membranes were found to range from 10-100 microgram/ml.
Document ID
19880033052
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
Authors
Knisley, Keith A.
(Texas Univ. Houston, TX, United States)
Rodkey, L. Scott
(Texas, University Houston, United States)
Date Acquired
August 13, 2013
Publication Date
January 1, 1986
Publication Information
Publication: Journal of Immunological Methods
Volume: 95
ISSN: 0022-1759
Subject Category
Life Sciences (General)
Accession Number
88A20279
Funding Number(s)
CONTRACT_GRANT: NAS9-17403
CONTRACT_GRANT: NIH-AI-20590
Distribution Limits
Public
Copyright
Other

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