Document Detail


Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill.
MedLine Citation:
PMID:  2470825     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The tetrazolium salt (MTT) method involving conversion of MTT to coloured formazan by cells serving as indirect measurements of cell growth/cell kill has been reported by several groups, although technical problems have been encountered. The present investigation was undertaken in order to delineate what laboratory variables have direct influence on the sensitivity and reproducibility of the method. The pH of the extraction buffer was of the utmost importance, since it was demonstrated that a pH greater than 5 would give rise to false signals. Furthermore, modifying the composition of the extraction buffer, all formazan dye grains were solubilised, totally. A direct comparison with published methods demonstrated that only the modified method would yield 100% higher signals without increasing the background. In contrast to previous reports, it was shown that phenol red does not interfere with the measurements and no washing steps are required since all ingredients can be added subsequently. Serum proteins at concentrations up to 25% have no influence on the result. All samples can be measured in an ELISA scanner at 570 nm with little intra-assay variation.
Authors:
M B Hansen; S E Nielsen; K Berg
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of immunological methods     Volume:  119     ISSN:  0022-1759     ISO Abbreviation:  J. Immunol. Methods     Publication Date:  1989 May 
Date Detail:
Created Date:  1989-07-03     Completed Date:  1989-07-03     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  1305440     Medline TA:  J Immunol Methods     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  203-10     Citation Subset:  IM    
Affiliation:
Institute of Medical Microbiology, University of Copenhagen, Denmark.
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MeSH Terms
Descriptor/Qualifier:
Animals
Buffers
Cell Count
Cell Division*
Cell Line
Cell Survival*
Coloring Agents
Formazans
Hydrogen-Ion Concentration
Kinetics
Mice
Reproducibility of Results
Spectrophotometry
Staining and Labeling* / methods
Tetrazolium Salts*
Thiazoles*
Tumor Cells, Cultured / metabolism,  pathology*
Chemical
Reg. No./Substance:
0/Buffers; 0/Coloring Agents; 0/Formazans; 0/Tetrazolium Salts; 0/Thiazoles; 298-93-1/thiazolyl blue

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


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