Time-based analysis of silver-stained proteins in acrylamide gels

Silver staining of proteins after PAGE often remains the method of choice in many laboratories. Nevertheless, it is known that quantification of protein levels is keenly restricted to a small range of protein concentrations leading to an over‐ or underestimation of protein amounts. To overcome this,...

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Bibliographic Details
Published in:Electrophoresis 2006-05, Vol.27 (10), p.1867-1873
Main Authors: Becher, Bertram, Knöfel, Ann-Kathrin, Peters, Jörg
Format: Article
Language:English
Subjects:
Animals
Cattle
Dynamic range
Electrophoresis, Gel, Two-Dimensional - methods
Electrophoresis, Gel, Two-Dimensional - statistics & numerical data
Image Processing, Computer-Assisted
Protein quantification
Proteins - analysis
Quantifiable silver staining
Serum Albumin, Bovine - analysis
Silver
Silver staining
Staining and Labeling
Time Factors
Time-based analysis
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Summary:Silver staining of proteins after PAGE often remains the method of choice in many laboratories. Nevertheless, it is known that quantification of protein levels is keenly restricted to a small range of protein concentrations leading to an over‐ or underestimation of protein amounts. To overcome this, a time‐based analysis method was developed to avoid the saturation effect of the silver‐staining reaction, thus resulting in an improved dynamic range of the gel image produced and therefore better quantification of proteins. Instead of the well‐known end‐point image analysis, gray intensities of time series images of a developing gel are determined and times until a threshold gray value is reached are calculated. These times are used to calculate a new grayscale image which can be analyzed using commercial image processing software.
ISSN:0173-0835
1522-2683
DOI:10.1002/elps.200500733