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Figure 2 | Journal of Translational Medicine

Figure 2

From: Capillary nano-immunoassays: advancing quantitative proteomics analysis, biomarker assessment, and molecular diagnostics

Figure 2

CNIA assay performance at a glance. a Assay reproducibility. Size-CNIA data gathered from a CNIA instrument operated by four different users over 4 days. Each run consisted of 11 capillaries analyzing an identical HeLa lysate with an ERK1 antibody using the standard protocol. Instrument software calculated ERK1 peak area in each capillary, and this was used to calculate average and %CV of peak signal (Courtesy of ProteinSimple). b A comparison with conventional Western blot and data quantitation. Prostate cancer LNCaP cells were treated with the indicated concentrations of PMA, bryostatin 1, or bryostatin 7 for 24 h. PKCβII and PKCδ were analyzed in total cell lysates by Western blot and size-CNIA using anti-PKCβII and anti-PKCδ antibodies. Top and middle panels show representative images of conventional Western blot and CNIA respectively. Levels of PKCβII and PKCδ were quantitated from the CNIA data and shown in the lower panels. β-actin signals were used as loading controls, and normalized values were expressed relative to that of the DMSO-treated cells. Values represent the mean ± SEM of three independent experiments [38]. c ERK isoform responses to PMA treatment detected by Charge-CNIA. RasGRP3 transfected LNCaP cells were treated with indicated concentrations of PMA for 30 min. ERK1 and ERK2 signals were analyzed with charge-CNIA using a pan-ERK antibody. Left panel shows the peak profile for the ERK signals, indicating ERK phosphorylation induced by PMA treatment. Right panel shows the dose response curve of ERK isoforms to PMA treatment calculated from the CNIA data. Values represent the mean ± SEM of four independent experiments.

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