Evaluation of membranes used for electroblotting of proteins for direct automated microsequencing.
Protein purification and characterisation have been age-old problems for the biochemist. A new era has arisen with the advent of one- and two-dimensional gel electrophoresis and high sensitivity automated protein microsequencing. These two tools along with electroblotting have made it possible to separate and analyse complex protein mixtures. We studied six different membranes compatible with Edman degradation chemistry to determine their efficiencies at binding proteins electroblotted from one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Their overall blotting-sequencing properties were also evaluated. We found that the polyvinylidene difluoride-based membranes out-performed the glass-based and polypropylene-based membranes under our selected experimental conditions. The problems associated with electroblotting and microsequencing are discussed.
Department of Cardiothoraric Surgery, National Heart and Lung Institute, London, UK.
C S
CS
Baker
M J
MJ
Dunn
M H
MH
Yacoub
D002021:Buffers; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel; D005898:Glass; D008567:Membranes, Artificial; D011126:Polypropylenes; D011145:Polyvinyls; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification*; D005465:Fluorocarbon Polymers
D002021:Buffers; D003101:Collodion; D008567:Membranes, Artificial; D011126:Polypropylenes; D011145:Polyvinyls; D011506:Proteins; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers