Document Detail

Semidry electroblotting of peptides and proteins from acid-urea polyacrylamide gels.
MedLine Citation:
PMID:  9367507     Owner:  NLM     Status:  MEDLINE    
A semidry electrophoretic transfer method was developed for efficient electroblotting of proteins separated by acid-urea polyacrylamide gel electrophoresis (AU-PAGE). Model polypeptides ranging from 1.8 to 21.5 kDa were used to test transfer parameters that included time of transfer, power settings, transfer solutions, and membrane type. Optimized conditions were identified which allowed for transfer efficiencies of 70-100% following 5-15 min of applied current. The best transfer solution was 5% acetic acid, the same solvent used for electrophoresis. Therefore, acid-urea gels could be subjected to electrophoretic transfer without a soaking step, thereby reducing loss of band resolution and eliminating leaching of protein from the gel. The method was shown to be applicable to Western blot analysis of rat neutrophil defensins.
M S Wang; J S Pang; M E Selsted
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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:  253     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1997 Nov 
Date Detail:
Created Date:  1997-12-16     Completed Date:  1997-12-16     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  225-30     Citation Subset:  IM    
Copyright Information:
Copyright 1997 Academic Press.
Department of Pathology, College of Medicine, University of California, Irvine, California 92697, USA.
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MeSH Terms
Acetic Acid
Blood Proteins / analysis
Blotting, Western / methods*
Electrophoresis, Polyacrylamide Gel / methods
Membranes, Artificial
Neutrophils / chemistry
Peptides / analysis*
Proteins / analysis*
Rats, Sprague-Dawley
Grant Support
Reg. No./Substance:
0/Blood Proteins; 0/Defensins; 0/Gels; 0/Membranes, Artificial; 0/Peptides; 0/Polyvinyls; 0/Proteins; 24937-79-9/polyvinylidene fluoride; 57-13-6/Urea; 64-19-7/Acetic Acid; 9004-70-0/Collodion

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