1/21/2024 0 Comments Licor western blot analysisA library of ~2500 known bioactive compounds was used. A pilot small molecule siRNA screen was performed for inhibitors of mTORC1 signaling. Replicability of In-Cell Western Assay for siRNA screening via a correlation plot. B) Signal intensities from In-Cell Western wells are plotted as a proportion of the mean values for each of two plates (plate 1 in blue, plate 2 in red). Band intensities are expressed as a proportion of the mean of the thirteen samples on each blot (blot 1 in blue, blot 2 in red). A) GAPDH and PMLC20 were detected on duplicate Western blots (13 replicates per blot, not shown). Intra-assay variability of Western blots and In-Cell Western Assays. In-Cell Western Assays offered superior precision, reduced variability, and smaller CVs. In-Cell Western Assay and Western blot analysis yielded very similar results (Figure 1). 1 Primary cultures of uterine myocytes stimulated with oxytocin were used to assess specificity, sensitivity, and precision of the two methods for phospho-analysis. 2, 4, 5Ī 2010 study compared In-Cell Western Assays and Western blotting (WB) for measurement of phosphorylated myosin regulatory light chain (PMLC20). If they overlap, or nearly overlap (Z′ 0.5). Z′ is calculated by running a large number of positive and negative controls and determining how much separation there is between positive and negative controls. Z′-Factor can provide some indication of the replicability of an assay. Z′-Factor is a measure of statistical effect size and can be used to assess if a response to an assay requires further investigation.
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