Document Detail


Monoclonal antibody to calmodulin: development, characterization, and comparison with polyclonal anti-calmodulin antibodies.
MedLine Citation:
PMID:  1862939     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Specific anti-calmodulin rabbit polyclonal and murine monoclonal antibodies have been produced with a thyroglobulin-linked peptide corresponding to amino acids 128-148 of bovine brain calmodulin. The monoclonal antibody is IgG-1 with kappa light chains. Both sets of antibodies recognize native vertebrate calmodulin, with the polyclonal antibody exhibiting an approximately fourfold higher sensitivity than the monoclonal antibody in a radioimmunoassay. The affinity of both polyclonal and monoclonal antibodies is approximately 2.5-fold higher for Ca(2+)-free calmodulin than for Ca(2+)-calmodulin. Other selected members of the calmodulin family (S100, troponin, and parvalbumin) do not exhibit significant cross-reactivity with the monoclonal antibody. Troponin and S100 beta displace some 125I-calmodulin from the polyclonal antibody, but require at least 900-fold excess concentration. The monoclonal antibody recognizes intact vertebrate calmodulin in solution and also on solid-phase. In addition, plant calmodulin and some forms of post-translationally modified calmodulin (phosphorylated or glycated) bind the monoclonal antibody. The affinity of the monoclonal antibody is approximately 5 x 10(8) liters/mol determined by displacement of 125I-calmodulin. On dot blotting the sensitivity for vertebrate calmodulin is 50 pg. The epitope for the monoclonal antibody is in the carboxyl terminal region (residues 107-148) of calmodulin. This highly specific anti-calmodulin monoclonal antibody should be a useful reagent in elucidating the mechanism by which calmodulin regulates intracellular metabolism.
Authors:
D B Sacks; S E Porter; J H Ladenson; J M McDonald
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Analytical biochemistry     Volume:  194     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1991 May 
Date Detail:
Created Date:  1991-08-30     Completed Date:  1991-08-30     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  369-77     Citation Subset:  IM    
Affiliation:
Department of Pathology, Brigham & Women's Hospital, Boston, Massachusetts.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibodies / immunology*
Antibodies, Monoclonal / biosynthesis,  immunology*
Antibody Specificity
Calcium / metabolism
Calmodulin / immunology*,  metabolism
Cattle
Protein Processing, Post-Translational
Rabbits
Radioimmunoassay
Grant Support
ID/Acronym/Agency:
DK01680/DK/NIDDK NIH HHS; DK25897/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Antibodies; 0/Antibodies, Monoclonal; 0/Calmodulin; 7440-70-2/Calcium

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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