Product uses
- Substrate for activation of kinesin ATPases (see Fig. 1)
- Determination of IC50s for kinesin inhibitors
- Used in Cytoskeleton, Inc’s kinesin ELIPA and End point ATPase assays (Cat. # BK060 & BK053)
- Substrate for discovery and characterization of microtubule binding proteins (MAPs) (see Fig. 2)
- Ideal for HTS applications
Material
Stabilized microtubules (MTs) are supplied as a lyophilized powder. Microtubules have been prepared from porcine brain tubulin protein that is greater than 99% pure (Cat. # T240). These stringently quality controlled MTs provide highly reliable and reproducible results in assays that require MT substrates. The average MT length in this product is 2 µm. Using these pre-formed microtubules in assays depending on microtubules gives very reproducible results. In a microtubule activated kinesin ATPase assay (see Fig. 1), the CV between samples (Vmax) was less than 5% and the CV of Vmax between assays using different batches of MT001 was less than 8%.
Purity
The tubulin protein used to make pre-formed microtubules is >99% pure. See Cat. # T240.
Biological Activity
MTs serve as a substrate for kinesin motor proteins. Kinesin motor proteins orchestrate a wide range of kinetic events within a cell. They have been shown to move cargoes such as chromosomes and vesicles along MT tracks. Kinesins operate by utilizing the energy of ATP to hydrolysis, an activity that is greatly enhanced in the presence of MTs. An MT activated Enzyme Linked Inorganic Phosphate ATPase (ELIPA™) assay (Cat. # BK060) can therefore be used as a test for the biological activity of MT001. The motor protein used in this assay is Kinesin Heavy Chain motor domain (KHC: Cat. # KR01). MT001 is also an excellent substrate for detecting microtubule binding proteins
