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Figure 6 | Malaria Journal

Figure 6

From: Development of two novel high-throughput assays to quantify ubiquitylated proteins in cell lysates: application to screening of new anti-malarials

Figure 6

Assay robustness and validation. A. Correlation of two plates with proteasome inhibitors located in known positions at four different concentrations simulating a screening plate (see embedded Table). Response produced by proteasome inhibitors on the levels of ubiquitylated proteins represented as % of stimulation (% of stimulation = 100*(sample/average control 1). The size of the points is proportional to the compound concentration tested. Light grey is the linear regression of the data while dark grey is the perfect correlation (y = x). Black dotted lines represent the statistical cut-off of each replicate (average control 1 + 3*standard deviation). Positive hits are above the cut off. B. Graph of control 1 (iRBCs without treatment), and control 2 (iRBCs with proteasome inhibitor 1.5 μM MG132 or 15 μM lactacystin) in the screening plates (n = 96 wells) represented as % of stimulation. C. Comparison of AlphaLISA (purple) and DELFIA (black) assay control data: robust average in bars, robust CV in italics and S/B in bold for control 1, control 2 and control without lysates in both assays.

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Malaria Journal

ISSN: 1475-2875

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