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Tubulin Polymerization AssayBiochem Kit: soybean tubulin

Manufacturer: Cytoskeleton Inc
Shipping: Standard

Product Uses Include :

  • Screen for dicotyledon targeted herbicides.
  • Basic research to measure effects of proteins and compounds on polymerization activity.
  • Basic research plant tubulin polymerization studies.
  • Source of plant tubulin for monomer or polymer (microtubule) binding studies (similar to BK029).
     

Introduction :
This assay is a one step procedure for performing plant tubulin polymerization assays. Polymerization is followed by fluorescence enhancement due to the incorporation of a fluorescent reporter into microtubules as polymerization occurs. It is an adaptation of an assay originally described by Bonne, D. et al. (1) for bovine brain tubulin. By calculating Vmax of polymerization in AFU/min you can determine the specific activity of a compound or protein on plant tubulin polymerization.

Example results :
Compounds or proteins that interact with tubulin will often alter one or more of the characteristic phases of polymerization.  For example, Figure 1 shows the effect of adding the anti-mitotic drug paclitaxel to the standard polymerization reaction.  A 3 µM concentration of paclitaxel eliminates the nucleation phase and enhances the Vmax of the growth phase.  Thus, one application of this assay is the identification of novel anti-mitotic compounds that enhance polymerization and stabilize microtubules.  Figure 1 also shows the effect of adding the tubulin binding drug, trifluran.  At 3 µM final concentration, trifluran causes a drastic decrease in Vmax and reduction in final polymer mass. 

Kit contents :
The kit contains sufficient material for 24 assays in 10 µl format. The following components are included:

  1. >90% pure tubulin (Cat. # TP005)
  2. General tubulin buffer with fluorescent reporter
  3. Compound dilution buffer
  4. GTP solution (Cat. # BST06)
  5. Paclitaxel positive control (Cat. # TXD01)
  6. Small volume 384-well plate.
  7. Manual with detailed protocols and extensive trouble shooting guide
     

Equipment needed :

  1. Tempererature controlled 384-well plate fluorimeter equipped with filters for exitation at 340-360 nm and emission at 420-450 nm

References :

Bonne, D., Heusele, C., Simon, C., and Pantaloni, D.  (1985).  4’, 6-Diamidino-2-phenylindole, a fluorescent probe for tubulin and mictrotubules. J. Biol. Chem. 260, 2819-2825.

SizeCatalogue noPrice 
30-36 Assays Kit size BK010S £1,295.00/€1,813.00
Qty:

All prices shown are exlusive of VAT.