Electroimmunoblotting of small peptides separated on urea-dodecyl sulphate (SUDS) gels. Application to myelin basic protein.
A method for the electroimmunoblotting and immunodetection of peptides of less than 50 amino acid residues is described. Excellent resolution of a mixture of myelin basic protein (MBP) peptides was achieved by electrophoresis in a polyacrylamide stacking, urea-dodecyl sulphate minislab gel. Following electrophoresis, the peptides were transferred to various matrices and probed with monoclonal and polyclonal antibodies. Variables such as transfer time, membrane type, fixation and the amount of peptide loaded on the gel have been optimized as a consequence native and synthetic peptides can now be visualized in gels and immunodetected on immobilizing matrices. This procedure is particularly suited to the analysis and identification of small MBP fragments arising in various neuropathological conditions as well as for the rapid characterization of antigenic determinants recognized by monoclonal and polyclonal anti-MBP antibodies.
D000818:Animals; D002417:Cattle; D003101:Collodion; D004586:Electrophoresis / Q000379:methods*; D007163:Immunosorbent Techniques; D008970:Molecular Weight; D004676:Myelin Basic Protein / Q000032:analysis*; D009757:Nylons; D010455:Peptides / Q000032:analysis*; D011817:Rabbits; D014508:Urea
D004676:Myelin Basic Protein; D009757:Nylons; D010455:Peptides; D014508:Urea; D003101:Collodion