微管蛋白聚合试剂盒cytoskeleton公司

Product Uses Include

• Screening compounds for effects on tubulin polymerization activity.
• Screening proteins for effects on tubulin polymerization activity.
• Teaching aid for undergraduate/graduate class in pharmacology.
• Economical approach to HTS with tubulin.
   

If you are interested in using either of these tubulin polymerization assays in a high throughput setting, please contact our technical service department for advice and bulk quotes.
 

Kit contents
This kit contains enough materials for 24 assays (BK004P). The following reagents are included:

1. HTS tubulin, lyophilized 
2. GTP solution (Cat. # 
3. General tubulin buffer (PEM, Cat. 
4. Tubulin glycerol buffer (Cat. 
5. Paclitaxel positive control (Cat. #
6. DMSO for paclitaxel.
7. Half area 96-well plate for polymerization reactions
8. Manual with detailed protocols and extensive troubleshooting guide.
    

Equipment needed

1. 96-well plate spectrophotometer with filters to read optical density at 340 nm.
2. Multi-channel pipette for rapid pipetting of tubulin
    

Example results
The BK004P kit was used to study the effect of 48 different conditions on tubulin polymerization rates in a 96 well format. Each condition was tested in duplicate samples (Fig. 1)

	Figure 1. Tubulin polymerization in 96-well format using BK004P. 48 different tubulin polymerization conditions were tested in duplicates. Wells A1 and A2 represent a control reaction and wells C1 and C2 represent the addition of the microtubule stabilizing drug Paclitaxel, resulting in an increased polymerization rate. Each well represents one polymerization reaction. The x-axis is 20 min and the y-axis corresponds to 0.0-0.2 OD at 340 nm.

References

1. Shelanski, M. L., Gaskin, F. and Cantor, C. R. (1973). Microtubule assembly in the absence of added nucleotides. Proc. Natl. Acad. Sci. U.S.A. 70, 765-768.
2. Lee, J. C. and Timasheff, S. N. (1977). In vitro reconstitution of calf brain microtubules: effects of solution variable. Biochemistry, 16, 1754-1762.

Lamberth et al., 2012. Synthesis and fungicidal activity of tubulin polymerisation promoters. Part 2: Pyridazines. Bioorg. Med. Chem. v 20, pp 2803-2810.

Kamal et al., 2012. 3-Substituted 2-Phenylimidazo[2,1-b]benzothiazoles: Synthesis, Anticancer Activity, and Inhibition of Tubulin Polymerization. ChemMedChem. v 7, pp 292-300.

Ahn et al., 2011. A novel bis-indole destabilizes microtubules and displays potent in vitro and in vivo antitumor activity in prostate cancer. Cancer Chemother. Pharmacol. v 67, pp 293-304.

Butler et al., 2011. Neurodegenerative Effects of Recombinant HIV-1 Tat(1-86) are Associated with Inhibition of Microtubule Formation and Oxidative Stress-Related Reductions in Microtubule-Associated Protein-2(a,b). Neurochem. Res. v 36, pp 819-828.

Lee et al., 2011. Biological evaluation of KRIBB3 analogs as a microtubule polymerization inhibitor. Bioorg. Med. Chem. Lett. v 21, pp 977-979.