PMID: 3760586Sep 27, 1986Paper

Computer-assisted determination of protein concentrations from dye-binding and bicinchoninic acid protein assays performed in microtiter plates

Journal of Immunological Methods
R D LaneB L Beck

Abstract

The bicinchoninic acid protein assay was scaled down to operate in the wells of microtiter plates. The total volume of the assay was reduced to 210 microliters and required only 10 microliters of sample per assay. The assay can be rapidly performed and then read with a plate reader. The analysis of the data was performed with a microcomputer interfaced with the reader. A computer program was developed to analyze the data from the bicinchoninic acid assay as well as the dye-binding assay at either the high or low protein concentration ranges. Based upon the absorbance values of the standards, the program computed a linear regression formula which was then used to calculate the concentrations of the sample proteins. A series of assays performed using bovine serum albumin, alpha-chymotrypsin and gamma globulin demonstrated that the scaled down bicinchoninic acid assay produced linear absorption versus concentration data with average correlation coefficients between 0.9967 and 0.9940 at the high and low concentration ranges respectively. The protein-to-protein variation and proper selection of either protein assay based upon the presence of interfering materials was considered.

References

May 1, 1977·Analytical Biochemistry·J J Sedmak, S E Grossberg
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Jan 28, 1983·Journal of Immunological Methods·M D RichardsonP A Llewellyn
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Nov 1, 1983·Analytical Chemistry

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Citations

Feb 3, 2007·Pediatric Nephrology : Journal of the International Pediatric Nephrology Association·Gerald SchwerdtMichael Gekle
Mar 28, 1995·Proceedings of the National Academy of Sciences of the United States of America·C S KimD L Alkon
Jun 1, 1990·International Journal of Andrology·E Töpfer-Petersen, D Cechová
Oct 13, 2006·Kidney & Blood Pressure Research·Andreas BenesicMichael Gekle
May 20, 1998·Proceedings of the National Academy of Sciences of the United States of America·A FavitD L Alkon
Jul 4, 2012·Free Radical Biology & Medicine·Stefanie RuhsClaudia Grossmann
Nov 27, 2008·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Gerald SchwerdtMichael Gekle
Jan 1, 1988·Biological Chemistry Hoppe-Seyler·E Töpfer-Petersen, A Henschen
Nov 1, 1993·Scandinavian Journal of Clinical and Laboratory Investigation·T OsnesP Kierulf
Mar 22, 2012·Journal of Vascular Research·Gerald SchwerdtMichael Gekle
Dec 21, 1987·FEBS Letters·E Töpfer-Petersen, A Henschen
Mar 18, 2006·Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology·Karina DrummMichael Gekle
Jan 6, 2016·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Maria BretschneiderClaudia Grossmann
Dec 29, 2017·Cellular and Molecular Life Sciences : CMLS·Mirjana PolovicMichael Gekle
Aug 16, 2019·Biochimica Et Biophysica Acta. Molecular Cell Research·Marie-Christin SchulzGerald Schwerdt
Feb 15, 1989·Analytical Biochemistry·R C Goldschmidt, H K Kimelberg

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