ELISA Kit Detects Murine Kras Activity with Ectopic Expression

August 24, 2022

Duke Univ

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Company:

Abcam

Product Name:

Ras GTPase ELISA Kit (Chemiluminescent)

Catalog Number:

ab134640

I wanted to quantitatively compare Raf1-RBD binding (GTPase activity) of different mutants of murine Kras constructs ectopically expressed in human HEK-HT cells. This kit delivered the result and demonstrated slight differences in Ras activity in different mutants. Note that the kit is only approved for human Hras & Kras; not for murine Kras. We overcame this obstacle by overexpressing murine Kras ectopically.

Experimental Design and Results Summary

Application

ELISA (chemiluminescence)

Starting Material

Cell lysate

Tips

Blank measurements and negative controls are important. If possible, opt for 3 technical replicates (reads).

Results Summary

This assay enabled the quantitative demonstration of slight differences in Ras activity of murine constructs that have 2 different mutations by overexpressing in human cells. However, the assay didn't detect any activity of murine Kras endogenously expressed in mouse cell lines or mouse tissues.

DOI or PMID #

N/A

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Summary

The Good

Quantitative measurement of RAS activity of human Hras and Kras.

The Bad

Not great for activity measurement of murine Kras when expressed endogenously.

The Bottom Line

The kit works great for human Kras. I wouldn't recommend it for endogenous murine Kras.

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