Document Detail


Enhancement of enzyme-activated 1,2-dioxetane chemiluminescence in membrane-based assays.
MedLine Citation:
PMID:  7710059     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Processes for enhancing the chemiluminescent signal generated by enzyme-activated 1,2-dioxetanes have been developed for membrane-based assays in which detection is done using a charge-coupled device (CCD) camera or X-ray film. The enhancement is demonstrated using slot-blots of biotinylated lambda DNA in conjunction with an avidin-alkaline phosphatase conjugate. For detection with a CCD camera, the nylon membrane is dried after processing and incubation in dioxetane substrate solution and heated to temperatures of 50 to 80 degrees C during detection. Up to a 100-fold signal increase is obtained using this enhancement process compared to the conventional detection procedure, in which the blot is kept saturated with substrate solution in a sealed plastic bag during detection. For detection with X-ray film, a fivefold increase in signal intensity is realized by drying the membrane before exposure to the film. These enhancement processes greatly reduce the time required for detection in membrane-based assays.
Authors:
R K Kobos; B A Blue; C W Robertson; L A Kielhorn
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Analytical biochemistry     Volume:  224     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1995 Jan 
Date Detail:
Created Date:  1995-05-10     Completed Date:  1995-05-10     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  128-33     Citation Subset:  IM    
Affiliation:
Corporate New Business Development Division, DuPont Company, Incorporated, Wilmington, Delaware 19880.
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MeSH Terms
Descriptor/Qualifier:
Chemiluminescent Measurements
Heterocyclic Compounds / chemistry*
Signal Processing, Computer-Assisted
Temperature
X-Ray Film
Chemical
Reg. No./Substance:
0/Heterocyclic Compounds; 6788-84-7/1,2-dioxetane

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


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