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Accurate Measurement for Improving Yield
Do your cells express the developed target protein? Do your culture supplements raise product titers? Is cell culture scaling affecting protein yield? It’s sometimes difficult to quantify protein expression when you have to measure your tailored protein in a complex sample matrix using traditional optical detection technologies. Now you can use your antibodies with an AMMP® non-optical detection assay and the ViBE® workstation to rapidly convert your current assays to work in challenging samples.
The versatile ViBE® utilizes BioScale’s proprietary AMMP® technology, a non-optical detection technique that offers protein researchers an easy to use platform that uniquely addresses protein analysis needs with highly sensitive detection and quantitation in complex biological mixtures.
See how BioScale can help you achieve your research goals. To discuss your biomarker or protein analysis application contact us.
Monitoring Protein Expression in Xenograft Tumor Lysates
Sample effects can make developing and using traditional detection schemes difficult and sometimes impossible on critical time lines. Gadd34 or Growth Arrest and DNA Damage protein 34 is a stress-induced protein implicated in the control of protein synthesis and apoptosis (1). Under normal non-stressed conditions Gadd34 is a rapidly degraded protein through the ubiquitin-proteasome pathway(2). Because proteasome inhibition by bortezomib or its analogs induces Gadd34 accumulation Millenium Pharmaceuticals sought to develop a strategy for monitoring increases in protein levels. In their study, a complementary pair of rabbit monoclonal antibodies was developed against recombinant Gadd34. These antibodies were then affinity tagged and used to generate a standard curve with recombinant Gadd34. Cell culture derived HCT116 or CWR22 xenograft tumor lysates either treated or untreated with proteasome inhibitors were analyzed on the ViBE® Bioanalyzer and compared to results obtained with either LiCor® Western Blot or AlphaScreen™. They concluded that the use of the ViBE® eliminates the labor intensive effort of Western Blot analysis and, furthermore, is devoid of the optical and chemical interferences derived from lysates of xenograft tumors observed with AlphaScreen™.
Induction of GADD34
Induction of Gadd34 of HCT116 derived cell culture lysates in the presence or absence of boronate proteasome inhibitor as analyzed by either ViBE® BioAnalyzer (Panel A) or AlphaScreen™(Panel B) or Western blot LiCor® (Panel C). HCT116 cells were cultured in the presence or absence of 3 nM of a bortezomib analog and lysed in mPER buffer at the times indicated. Protein concentrations were determined by BCA and normalized for sample addition for each assay. Results shown are representative of at least duplicate experiments with standard errors of less than 10%.