Excellent Kit For Measuring Kinesin Activity

January 10, 2018

University of Kansas Medical Center
Pathology
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cytoskeleton

Product Name:

HTS kinesin ATPase Endpoint Assay Biochem Kit

Catalog Number:

BK053

We are interested in kinesins proteins' roles in tumorigenesis and drug resistance. We have generated recombinant kinesin proteins and used this assay to determine whether the proteins are enzymatically active.

Experimental Design and Results Summary

Application

ATPase Biochemical Assay

Starting Material

Recombinant proteins

Protocol Overview

To determine specificity of the assay for ADP, we measured the assay sensitivity for inorganic phosphate to generate a phosphate standard curve. Pi-free water and increasing amounts of phosphate (0.3 - 2.0 nmoles) were added to wells in duplicate, then the CytoPhos supplied with the kit was added. Plate was read on a Tecan spectrophotometer at 650 nm.

Tips

This kit works best with half-area 96-well plates, be sure to review the protocol and order additional supplies before you begin.

Results Summary

Excellent data with high R values for the curve. Reproducible over years of use, with multiple scientists performing assays.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

This is a very fast and used friendly assays for measuring production of Pi.

The Bad

The kit is costly but, it lasts longer than the 1 year shelf life.

The Bottom Line

Great purchase to quickly design an assay for measuring inorganic phosphate production as a measure of kinesin motor activity.

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