An automated Western blot analysis using the phastsystem.
We have developed a rapid, automated method for preparing Western blots of very small amounts of proteins, utilizing a commercially available electrophoresis system (Phastsystem, Pharmacia). This system has been adapted to transfer to nitrocellulose experimental samples that were initially separated in the same system by gradient-sodium dodecyl sulfate gel electrophoresis. The developing unit of the Phastsystem has permitted automation of all the necessary steps including incubation with antibodies, saturation of nonspecific binding sites, and washing. The total elapsed time for protein separation, transfer, and staining is about 6 to 7 h.
Prieur
B
B
Unité Associée Institut Pasteur/INSERM No. 285, Institut Pasteur, Paris, France.
Russo-Marie
F
F
D000818:Animals; D017302:Annexins; D001322:Autoanalysis; D001345:Autoradiography; D015153:Blotting, Western / Q000295:instrumentation / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000302:isolation & purification; D007136:Immunoglobulins / Q000302:isolation & purification; D011817:Rabbits
D017302:Annexins; D006023:Glycoproteins; D007136:Immunoglobulins