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Detection of Phosphorylated and Total ERK and p38 MAPK by Bio-Plex Assay, ELISA, and Western Blotting, Rev A

well and 1 g/well; the reported sample absorbance reached a plateau, and the highest OD reported was only 1.755 (Figure 4A). The Bio-Plex assay demonstrated distinctly separate signals across all the samples (Figure 4B), which correlated well with the western blot bands (Figure 4C).


Conclusions
For the detection of total p38 MAPK in HEK 293 cells, both the Bio-Plex assay and ELISA correlated well with the western blot. The Bio-Plex assay showed saturation at the high end but potential to detect the target in further-diluted samples. With the HeLa lysate, the Bio-Plex assay detected the target with good sensitivity and demonstrated a broad detection range that correlated well with the western blot bands. The target was not detectable by ELISA

For the detection of phospho-p38 MAPK in HEK 293 cells, the Bio-Plex assay demonstrated a broad detection range at the high end and good correlation with the western blot. Bio-Plex and ELISA assays both showed potential for higher sensitivity toward more dilute samples

For the detection of total ERK, the Bio-Plex assay demonstrated better signal distinction and sensitivity compared to ELISA

For phospho-ERK detection, the ELISA showed saturation at the high end. The Bio-Plex assay showed better signal distinction, dose response, and correlation with western blots compared to ELISA


The Bio-Plex suspension array system includes fluorescently labeled microspheres and instrumentation licensed to Bio-Rad Laboratories, Inc. by the Luminex Corporation.

Information in this tech note was current as of the date of writing (2004) and not necessarily the date this version (Rev A, 2004) was published.
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