Improved dynamic range of protein quantification in silver-stained gels by modelling gel images over time.
Silver staining is a commonly used protein stain to visualise proteins separated by 2-DE. Despite this, the technique suffers from a limited dynamic range, making the simultaneous quantification of high- and low-abundant proteins difficult. In this paper we take advantage of the fact that silver staining is not an end-point stain by photographing the gels during development. This procedure provides information about the change in measured absorbance for each pixel in the protein spots on the gel. The maximum rate of change was found to be correlated with the amount of applied protein, providing a new way of estimating protein amount in 2-DE gels. We observed an improvement in the dynamic range of silver staining by up to two orders of magnitude.
Grove
Harald
H
Nofima Mat, Norwegian Institute of Food, Fisheries and Aquaculture Research, As, Norway. Harald.Grove@umb.no
Faergestad
Ellen M
EM
Hollung
Kristin
K
Martens
Harald
H
D000818:Animals; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005782:Gels / Q000737:chemistry; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D016014:Linear Models; D009124:Muscle Proteins / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D012015:Reference Standards; D012710:Serum Albumin, Bovine / Q000032:analysis; D016622:Silver Staining / Q000379:methods*
D005782:Gels; D009124:Muscle Proteins; D011506:Proteins; D012710:Serum Albumin, Bovine