Bromophenol Blue → Acrylamide

Participants

Reaction Steps

Standard cylinders of 10% acrylamide gel (9.7% acrylamide and 0.3% N,N'-methylene-bis-acrylogo there merized with TEMED) and ammonium persulfate were prepared and set up between gel and reservoir buffers comprising tris hydrochloride at pH 8.0 and 0.1% SDS. The upper gel surface was loaded with a charge comprising 50 micrograms of T2 pilus crystals, 20 ug of Clelands Reagent (0.01M) 1 mg. of SDS 20λ of glycol and 20λ of Bromophenol Blue (0.002%). Prior to charge the pili were heated with the SDS and the Cleland's reagent for 2 minutes at 100° C. The electrophoresis was run at 5 ma (at ca. 170 v) until the Bromophenol Blue had run 6 cm. The gels were removed and cut thru the dye band and two gels stained with Coomassie Blue Stain (0.2%) to give two bands--a major band and a minor band.

Provenance & Evidence