By John K. Buolamwini, Alex A. Adjei
This booklet not just surveys the total enviornment of novel anticancer drug objectives, but additionally provides a wide-ranging number of state of the art concepts at present being utilized all through novel antitumor drug discovery and improvement. The tools are utilized to experiments related to such themes as immunotherapy, angiogenesis, melanoma metastasis, the telephone cycle, sign transduction inhibitors, apoptosis, antibodies, antisense molecules, microarray gene expression, move cytometry, and puppy imaging for melanoma objective validation. There also are confirmed tools for the preclinical identity of drug goals and for objective validation in the course of the scientific trials of novel brokers.
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Extra resources for Novel Anticancer Drug Protocols
17. 18. 19. 20. 21. 22. 23. 24. 5-mL microfuge tubes. 15-mL centrifuge tubes. 50-mL centrifuge tubes. Cell scraper. 100-mm cell culture plates. 5-mL polystyrene round-bottom tube. Deionized water. Absolute ethanol. 4. β-Mercaptoethanol. Dimethyl Sulfoxide (DMSO). 1% Ponceau Solution in 5% acetic acid). Tween 20 (Sigma). 4. 5 M. Bromodeoxy uridine (BrdU) 50 mM stock in water, heat at approx 37°C to get into solution. RNase A (DNase free). Propidium Iodide (50 µg/mL in PBS). Gammabind G Sepharose (Amersham).
Rad. Oncol. 11, 316–327. CH04,49-56,8pgs 55 02/05/03, 11:52 AM 56 Sarkaria 11. , Berlinck, R. G. , et al. (1998) High-throughput assay for G(2) checkpoint inhibitors and identification of the structurally novel compound isogranulatimide. Cancer Res. 58, 5701–5706. 12. , Lim, L. , Daly, J. , Li, A. , Jacobson, K. , and Roberge, M. (2000) Structure-activity relationships for G2 checkpoint inhibition by caffeine analogs. Int. J. Oncol. 16, 971–978. 13. Tibbetts, R. , Brumbaugh, K. , Williams, J. , et al.
Dry gel with a gel dryer for 45 min to 1 h at 80°C. 8. Expose film to autoradiography film and/or PhosphorImager screen. 9. Develop film in film developer and/or phosphorimager (see Fig. 1). 2. Measuring CDK Activity in Intact Cells by Using Phospho-Specific Antibodies Against Endogenous CDK Substrates If a chemical compound inhibits cdks in vitro, then it will be necessary to determine the phosphorylation status of the CDK substrate. Thus, either direct or indirect small molecule cdk inhibitors should lead to loss in the phosphorylation of endogenous substrates such as Rb, vimentin, NPAT, or p70S6 kinase (10–14) (see Fig.
Novel Anticancer Drug Protocols by John K. Buolamwini, Alex A. Adjei