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Kinesin Motility Kit

Introduction

This is a Fluorescence based motility assay.  To perform this assay effectively you will require a video microscope system with a shutter mechanism for time lapse image acquisition. Kinesin or dynein motors are coupled to the surface of a mcroscope perfusion chamber (kinesin heavy chain motor domain is included in the kit as a positive control) and fluorescent microtubules are added. The motor-induced motility of the microtubules can be monitored and measured in a fluorescence microscope.
 

Kit contents

The kit contains sufficient materials for 25 assays. The following components are included:

   1. Tubulin, >99% pure, lyophilized (Cat nr TL238)
   2. Rhodamine tubulin, lyophilized (Cat nr TL331M)
   3. Kinesin heavy chain motor domain, lyophilized (positive control) (Cat nr KR01)
   4. Antifade
   5. ATP (Cat nr BSA04)
   6. Microtubule polymerization buffer
   7. Tubulin glycerol buffer
   8. Microtubule resuspension buffer
   9. Blocking solution
  10. Chamber wash buffer
  11. Motility buffer
  12. Paclitaxel (Cat nr TXD01)
  13. DMSO
  14. 25 acid washed perfusion chambers
  15. Chamber filters
  16. Manual with detailed protocols and extensive troubleshooting guide

Equipment needed

Fluorescence microscope equipped with filters to detect rhodamine and ready for time-lapse image capturing

Example results

The motility of microtubules in a kinesin heavy chain (Cat nr KR01) coated chamber was followed with time lapse microscopy (Fig. 1)




Figure 1. Microtubule Motility Assay. The field was selected to show a sparse area of MT coverage in order to allow clear visualization of MT motility. Each frame represents a 5 min time-point, from 0 to 15 min. The position of each MT at time zero is marked by an arrow. The arrow position remains identical in each frame to serve as a reference point for MT movement. To obtain an average MT velocity, we recommend measure motility of approximately 30 - 50 individual MTs.

Product Uses Include

Examples of publications where this product was used