Two Ways to Get Separation, Identification and Quantitation
Peggy Sue performs Simple Western assays in two ways. She can separate proteins by size or she can separate them by charge. Either way, she does the entire process for you, up to 96 samples at a time with as little as 0.05 µg/mL protein. On top of it, she can help you identify your proteins and quantitate them with confidence.
Figure 1. Analyzing post-translational modifications of Histone H3 is no problem for Peggy Sue. Left: HeLa cell lysate from cells stimulated with sodium butyrate was probed with anti-acetyl Histone H3 (Lys9) to characterize changes in acetylation. Right: Methylation was analyzed in lysate from HeLa cells stimulated with colcemid and probed with anti-dimethyl Histone H3 (Lys39).
Peggy Sue provides two different perspectives for quantitating biological responses.
Figure 2. The same HeLa lysate sample, stimulated with epidermal growth factor, was analyzed and probed with an anti-phospho specific ERK antibody in both size-based and charge-based assays. The size-based electropherogram shows detection of total phosphorylated ERK2 (pERK2 + ppERK2), a quantifiable large peak; and total phosphorylated ERK1 (pERK1 + ppERK1), a smaller peak, at their correct molecular weights. The charge-based electropherogram provides more in-depth characterization of the single and dual phosphorylated isoforms of ERK1 and ERK2 at their expected pIs, demonstrating that the dual phosphorylated isoforms are the dominant species present in the lysate.