Quantification of post-translational modifications in correlation to total protein concentration is very easy with Simple Western Technology

Udo Burger, Proteinsimple, Bio-Techne, Wiesbaden, Germany (udo.burger@bio-techne.com)

Research on biological processes and disease states require information on cross-talking networks of receptors and intracellular protein components. Combined with fluctuating post-translational modifications calls for a sensitive, quantitative and reproducible analytical technique. In complex samples, the highly selective Western blot technique has established itself as the method of choice for life science researchers across the board. Fluctuations in protein modifications often correlate to other modifications and change relatively to loading controls or total protein concentration. Commonly, loading controls are seen at higher abundancy or in the same size MW range as protein targets, which complicates assay optimization.

With Simple Western technology, quantification of protein targets in complex matrices are established quickly to give highly reproducible and sensitive results. Jess, the newest member of the Simple Western family, meets even your most challenging multiplexing needs with additional detection channels.  In addition to chemiluminescence, the NIR and IR channels offer the detection of proteins with identical masses and varying abundancies.

In cell lysate, upregulation and phosphorylation of epidermal growth factor receptor (EGFR) could be demonstrated with antibodies against EGFR and p-EGFR. Both signals have been independently analyzed in the same sample using different detection channels.

In addition, the total protein concentration could be determined within the same capillary on the same sample. The normalization of the target protein area against the total protein concentration was performed automatically using the analysis software Compass for SW.

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Short Biography of Presenting Author

Dr. Udo Burger

Biologist and Chemist

2002-2011 Support for bio-analytical chromatography and LC-MS in different companies (Shimadzu, Thermo Fisher Scientific, Clinical Diagnostic laboratoriers). Since August 2015 Field Applications Scientist Europe at Proteinsimple (Brand of Bio-Techne)