Western blots of SDS-PAGE protein samples before (A) and after (B, C, and D) each enrichment step. Western blots were performed using GP1. Although the identity of the samples applied to each lane was the same as illustrated in Figure 3, the amount of protein applied to each lane differed. (A). Fraction Sp, 5 μg, (B). Galactose eluted protein from the RCA column containing L-Arg stimulated ion channel activity, 1.6 μg, (C). Protein from the first peak fractions of the gel filtration step containing L-Arg stimulated ion channel activity, 0.2 μg, (D). Protein fraction containing L-Arg stimulated ion channel activity from the pH 9 fraction of the ion exchange procedure, 0.2 μg. Note that in spite of lowering protein amounts in lanes A – D, the intensity of the Western blot increases, indicative of an enrichment of the antigen protein(s) near 82–84 kDa.
Grosvenor et al. BMC Neuroscience 2004 5:25 doi:10.1186/1471-2202-5-25