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Isoelectric focusing-affinity immunoblot analysis of mouse monoclonal antibodies to the four human IgG subclassesIsoelectric focusing (IEF)/affinity immunoblotting and enzyme-linked immunosorbent assay (ELISA) were used for parallel analysis of murine monoclonal antihuman IgG-subclass antisera (MoAbs). Coomassie Blue-stained protein bands in the pH region 5.5-8.0 were shown to be murine IgG by direct blotting onto nitrocellulose followed by detection with conjugated antimouse IgG. Use of IgG myeloma antigen-coated nitrocellulose in the IEF-affinity immunoblot allowed detection of the charge microheterogeneity of MoAbs. The MoAb group contained one to five major dense bands flanked by up to four minor fainter bands, all with pIs ranging from 6.1 to 7.8. Semiquantitative estimates of binding specificity in the IEF-affinity blot compared well with cross-reactivity data obtained from a quantitative ELISA.
Document ID
19880027669
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
External Source(s)
Authors
Hamilton, Robert G.
(Texas Univ. Houston, TX, United States)
Roebber, Marianne
(Texas Univ. Houston, TX, United States)
Rodkey, L. Scott
(Texas, University Houston, United States)
Reimer, Charles B.
(PHS, Centers for Disease Control, Atlanta GA, United States)
Date Acquired
August 13, 2013
Publication Date
March 1, 1987
Publication Information
Publication: Electrophoresis
Volume: 8
ISSN: 0173-0835
Subject Category
Inorganic And Physical Chemistry
Accession Number
88A14896
Funding Number(s)
CONTRACT_GRANT: NIH-AI-20590
CONTRACT_GRANT: NAS9-17403
Distribution Limits
Public
Copyright
Other

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