Protein blotting followed by microsequencing.
The use of new membranes such as activated or derivatized glass fibers as well as synthetic membranes, which are compatible with the hazardous sequencing reagents, are described. Precautions to be taken in order to prevent N-terminal blockage of the proteins during electrophoresis and blotting are described, as well as the conditions for protein detection after blotting and protein treatment for in situ amino acid analysis, fragmentation and microsequencing. For a number of standard proteins and bacterial ribosomal proteins microsequence analysis is reported for two commercially available sequencers (Applied Biosystems and Knauer).
Choli
T
T
Brigitte Wittmann-Liebold, Max-Planck Institut für Molekulare Genetik, Abteilung Wittmann, Berlin, Germany.
Wittmann-Liebold
B
B
D000595:Amino Acid Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D005898:Glass; D006801:Humans; D009124:Muscle Proteins / Q000737:chemistry / Q000378:metabolism; D011145:Polyvinyls; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D012269:Ribosomal Proteins / Q000737:chemistry / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate; D012995:Solubility; D005465:Fluorocarbon Polymers
D009124:Muscle Proteins; D011145:Polyvinyls; D011506:Proteins; D012269:Ribosomal Proteins; D012967:Sodium Dodecyl Sulfate; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers