Concentrating protein samples for sodium dodecyl sulphate-polyacrylamide gel electrophoresis and isoelectric focusing using protein-blotting membranes.
The protein concentration in biological samples is a crucial limiting factor for a successful analysis by electrophoresis. Many techniques have been adopted to increase protein concentrations, however, they are often insufficient and require special equipment or poisonous chemicals. Herein, we report a simple and efficient technique for concentrating dilute protein samples by absorbing proteins onto protein-blotting membrane strips. In this technique, blotting membrane strips were incubated in dilute protein solutions to capture proteins. For either sodium dodecyl sulphate-polyacrylamide gel electrophoresis or isoelectric focusing, the protein-absorbed membrane strips were directly loaded to the sample wells which contained a strong protein elution buffer and electrophoresis was performed under standard conditions.
Liang
F T
FT
Department of Veterinary Science, University of Kentucky, Lexington 40546, USA.
Granstrom
D E
DE
Shi
Y F
YF
D015153:Blotting, Western; D004220:Disulfides; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D007525:Isoelectric Focusing / Q000379:methods*; D009994:Osmolar Concentration; D011145:Polyvinyls; D012492:Salts; D012967:Sodium Dodecyl Sulfate; D005465:Fluorocarbon Polymers
D004220:Disulfides; D011145:Polyvinyls; D012492:Salts; D012967:Sodium Dodecyl Sulfate; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers