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Quantitative densitometry of proteins stained with coomassie blue using a Hewlett Packard scanjet scanner and Scanplot software.
Electrophoresis. 1997 Jan; 18(1):67-71.E

Abstract

In the present study we evaluated the performance of a software/scanner system that employed the Hewlett Packard (HP) ScanJet Plus and Scanplot Software for densitometric quantification of protein loads stained with Coomassie Brilliant Blue following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Gels with bovine serum albumin (BSA) standards, ranging from 0.125 to 10 micrograms, were scanned using reflectance densitometry with 127 microns step size in both the x and y directions and a resolution of 200 dots per inch. Densitometric volume was calculated for each protein band from scanner output in the tagged image file format (TIFF) by a customized software package, Scanplot V. 4.05 (Cunningham Engineering). Protein loads between 0.125 and 10.0 micrograms vs. volume were fit by a second-order regression: Volume = -0.58 x protein load2 + 16.82 x protein load + 7.87 (r = 0.991, p < 0.01). The same gels were scanned and quantified using a transmittance laser densitometer; densitometric volumes measured by both systems were highly correlated (r2 = 0.981, p < 0.01). Additional gels of BSA, smooth muscle myosin heavy chain (myosin), and actin displayed linear relationships between protein loads up to 4.0 micrograms and densitometric volume reflecting unique dye binding properties. We conclude that accurate and reproducible quantitative densitometry of SDS-PAGE can be performed using the HP ScanJet Plus scanner and Scanplot software.

Authors+Show Affiliations

Department of Physiology, Queen's University, Kingston, Ontario, Canada.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9059824

Citation

Vincent, S G., et al. "Quantitative Densitometry of Proteins Stained With Coomassie Blue Using a Hewlett Packard Scanjet Scanner and Scanplot Software." Electrophoresis, vol. 18, no. 1, 1997, pp. 67-71.
Vincent SG, Cunningham PR, Stephens NL, et al. Quantitative densitometry of proteins stained with coomassie blue using a Hewlett Packard scanjet scanner and Scanplot software. Electrophoresis. 1997;18(1):67-71.
Vincent, S. G., Cunningham, P. R., Stephens, N. L., Halayko, A. J., & Fisher, J. T. (1997). Quantitative densitometry of proteins stained with coomassie blue using a Hewlett Packard scanjet scanner and Scanplot software. Electrophoresis, 18(1), 67-71.
Vincent SG, et al. Quantitative Densitometry of Proteins Stained With Coomassie Blue Using a Hewlett Packard Scanjet Scanner and Scanplot Software. Electrophoresis. 1997;18(1):67-71. PubMed PMID: 9059824.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Quantitative densitometry of proteins stained with coomassie blue using a Hewlett Packard scanjet scanner and Scanplot software. AU - Vincent,S G, AU - Cunningham,P R, AU - Stephens,N L, AU - Halayko,A J, AU - Fisher,J T, PY - 1997/1/1/pubmed PY - 1997/1/1/medline PY - 1997/1/1/entrez SP - 67 EP - 71 JF - Electrophoresis JO - Electrophoresis VL - 18 IS - 1 N2 - In the present study we evaluated the performance of a software/scanner system that employed the Hewlett Packard (HP) ScanJet Plus and Scanplot Software for densitometric quantification of protein loads stained with Coomassie Brilliant Blue following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Gels with bovine serum albumin (BSA) standards, ranging from 0.125 to 10 micrograms, were scanned using reflectance densitometry with 127 microns step size in both the x and y directions and a resolution of 200 dots per inch. Densitometric volume was calculated for each protein band from scanner output in the tagged image file format (TIFF) by a customized software package, Scanplot V. 4.05 (Cunningham Engineering). Protein loads between 0.125 and 10.0 micrograms vs. volume were fit by a second-order regression: Volume = -0.58 x protein load2 + 16.82 x protein load + 7.87 (r = 0.991, p < 0.01). The same gels were scanned and quantified using a transmittance laser densitometer; densitometric volumes measured by both systems were highly correlated (r2 = 0.981, p < 0.01). Additional gels of BSA, smooth muscle myosin heavy chain (myosin), and actin displayed linear relationships between protein loads up to 4.0 micrograms and densitometric volume reflecting unique dye binding properties. We conclude that accurate and reproducible quantitative densitometry of SDS-PAGE can be performed using the HP ScanJet Plus scanner and Scanplot software. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/9059824/Quantitative_densitometry_of_proteins_stained_with_coomassie_blue_using_a_Hewlett_Packard_scanjet_scanner_and_Scanplot_software_ L2 - https://doi.org/10.1002/elps.1150180114 DB - PRIME DP - Unbound Medicine ER -