Product Uses Include
- Measuring microtubule-activated kinesin ATPase activity
- Discovering/characterizing kinesin inhibitors
- Discovering/characterizing kinesin regulating cofactors
Introduction
Eukaryotic kinesin motor proteins orchestrate a wide range of kinetic events within a cell. They have been shown to move cargo, such as chromosomes and vesicles, along microtubule tracks (1). They also play a major role in the organization of cytoskeletal architecture as evidenced in the establishment of the microtubule spindle during mitosis (2).
Kinesins operate by utilizing the energy of ATP hydrolysis to move along their microtubule (MT) substrates. Once a kinesin motor binds to its MT track, the ATPase rate of the motor is often enhanced several hundred to several thousand-fold (3). MT activated kinesin ATPase is a major parameter in motor function and serves as a powerful method to monitor and study kinesin activity under various experimental conditions.
As part of its Cytoskeleton Motor Werks (CMW) line of research reagents, Cytoskeleton, Inc. has developed the Kinesin ATPase End-Point Biochem Kit™ (BK053). The kinesin end-point assay is an extremely quick and economical way to measure inorganic phosphate (Pi) generated during the microtubule activated ATPase activity of kinesin motor proteins. Large numbers of assays can be performed simultaneously in a homogenous reaction, making the assay highly suitable for HTS applications. The assay is based upon a colorimetric change, measured at 650 nm.
More detailed kinetic studies for kinesin MT activated ATPase assays can be performed with Biochem Kit™ BK060. Cytoskeleton, Inc. also offers a wide range of kinesin motors.
Kit contents
The kit contains sufficient materials for 1000 assays. The following components are included:
- CytoPhos™ reagent
- Kinesin motor protein (Cat. # KR01)
- Pre-formed microtubules (Cat. # MT002)
- Kinesin Reaction Buffer
- Paclitaxel (Cat. # TXD01)
- Phosphate standard
- DMSO
- Manual with detailed protocols and extensive troubleshooting guide
Equipment needed
- Spectrophotometer capable of measuring absorbance at 650 nm wavelength.
- Small capacity (100-1000 µl) cuvettes or 96-well microtiter plates.
