We started using the Odyssey 7 or 8 years ago, I mean it really has changed the way we do certain experiments, and it has dramatically changed the reliability of how we interpret certain experiments.
Dr. Oliver Hantschel
Senior Post-Doctoral Researcher
Research Center for Molecular Medicine of the Austrian Academy of Sciences
Dr. Oliver Hantschel is a senior post-doctoral researcher at the Research Center for Molecular Medicine of the Austrian Academy of Sciences. In early 2011, he will start his own lab as an Assistant Professor, at the École Polytechnique Fédérale De Lausanne (EPFL) in Switzerland.
Dr Hantschel's research focuses on the pathophysiology and pathogenesis of leukemia in humans (CML). His group examines kinase mutations in mammalian cells and has a special interest in the BCR-ABL protein, which causes chronic myeloleukemia and some forms of B-cell acute lymphoblastic leukemia. Quantification is crucial for their signal transduction and structural biology studies. Dr Hantschel's group uses mass spectrometry and tandem-affinity purification to analyze protein complexes.
A Robust Solution for Quantitative Western Blotting
Dr Hantschel has used the Odyssey® Imager for 8 years, primarily for Western Blotting and In‑Cell Western™ assays. "It's quite important for me to get quantitative information about phosphorylation status of the proteins I'm working at, or to be able to quantify protein amounts in Western Blots from different experiments," says Hantschel.
Protein expression and phosphorylation of downstream signaling pathways are analyzed, and the Odyssey is a very important tool for analysis of mutant kinases. The expression levels of mutant proteins are measured, and immunoprecipitations are performed to examine interactions with other proteins.
This group is working to move their studies in a clinical direction.
"One of the main goals is to identify novel strategies to inhibit the BCR-ABL tyrosine kinase. BCR-ABL is a prototypic drug target and it has been studied for quite some time, and there are some very good drugs available," says Hantschel. "One of the major clinical problems is that quite a number of patients get resistant to these inhibitors. And so in order to offer good treatment for these resistant patients, it is necessary to exploit novel therapeutic strategies, and to provide inhibitors for these resistant forms."
Taking Guesswork Out of Western Blotting Analysis
With the Odyssey, we can reliably quantify. This has made quite a difference and is much better for us to rely on all results and things are much more reproducible now.
Prior to using the Odyssey, the group used ECL detection and film in their experiments. "Basically we had to just look at the bands and kind of estimate the ratio of expression and then for the second round of experiments we had to adjust again. It is known that ECL signal is dependent on the time of exposure, and can only be roughly used to adjust for differences in expression or activity."
"But with the Odyssey, we can reliably quantify. This has made quite a difference and is much better for us to rely on all results and things are much more reproducible now," says Hantschel. "We started using the Odyssey 7 or 8 years ago, I mean it really has changed the way we do certain experiments, and it has dramatically changed the reliability of how we interpret certain experiments."
We thank Dr Hantschel for his important work in cancer research, and are proud to consider him an "Odyssey Expert".
For more information about Dr. Hantschel, visit Research Centre for Molecular Medicine of the Austrian Academy of Sciences: CeMM.
Publications resulting from work on the Odyssey
- Hantschel, O., Nagar, B., Guettler, S., Kretzschmar, J., Dorey, K., Kuriyan, J., and Superti-Furga, G. (2003). A Myristoyl/Phosphotyrosine Switch Regulates c-Abl. Cell 112(6), 845-857. (IF: 31.2)
- Hantschel, O., Wiesner, S., Güttler, T., Mackereth, C. D., Remsing Rix, L.L., Mikes, Z., Dehne, J., Görlich, D., Sattler, M. and Superti-Furga, G. (2005). Structural basis for cytoskeletal association of Bcr-Abl/c-Abl. Mol. Cell, 19(4), 461-473. (IF: 14.6)
- Nagar, B., Hantschel, O., Seeliger, M., Davies, J.M., Weis, W. I., Superti-Furga, G. and Kuriyan, J. (2006). Organization of the SH3-SH2 Unit in Active and Inactive Forms of the c-Abl Tyrosine Kinase, Mol.Cell, 21, 787-798. (IF: 14.6)
- Bürckstümmer, T., Bennett, K. L., Preradovic, A., Schütze, G., Hantschel, O., Superti-Furga, G. and Bauch, A. (2006). An efficient tandem affinity purification procedure for the characterization of protein complexes in mammalian cells. Nature Methods, 3(12), 1013-1019. (IF: 16.9)
- Sherbenou, D.W., Hantschel, O., Turaga, L., Kaupe, I., Willis, S., Bumm, T., Press, R.D., Superti-Furga, G., Druker, B.J. and Deininger, M.W. (2008). Characterization of BCR-ABL deletion mutants from patients with chronic myeloid leukemia. Leukemia, 22(6), 1184-1190. (IF: 8.3)
- Filippakopoulos, P., Kofler, M., Hantschel, O., Gish, G.D., Grebien, F., Salah, E., Neudecker, P., Kay, L.E., Turk, B.E., Superti-Furga, G., Pawson, T. and Knapp, S. (2008). Structural coupling of SH2-kinase domains links Fes and Abl substrate recognition and kinase activation. Cell, 134(5), 793-803. (IF: 31.2)
- De Keersmaecker, K., Versele, M., Cools, J., Superti-Furga, G. and Hantschel, O. (2008). Intrinsic differences between the catalytic properties of the oncogenic NUP214-ABL1 and BCR-ABL1 fusion protein kinases. Leukemia, 22(12), 2208-2216. [corresponding author] (IF: 8.3)
- Wojcik, J., Hantschel, O., Grebien, F., Kaupe, I., Bennett, K.L., Barkinge, J., Jones, R.B., Koide, A., Superti-Furga, G. and Koide, S. (2010). A potent highly specific FN3 monobody inhibitor of the Abl SH2 domain. Nature Struct. Mol. Biol., 17(4), 519-527. (IF: 12.3)
- Sherbenou, D.W., Hantschel, O., Kaupe, I., Willis, S., Bumm, T., Turaga, L., Lange, T., Dao, K.H., Press, R.D., Superti-Furga, G., Druker, B.J. and Deininger, M.W. (2010). BCR-ABL SH3-SH2 domain mutations in chronic myeloid leukemia patients on imatinib, Blood, 116(17), 3278-3285. (IF: 10.6)
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