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Analysis of MEK1 by Differential Scanning Fluorimetry(DSF) (CAT#: STEM-MB-0772-WXH)

Introduction

MEK1 is a protein involved in a cell signaling pathway that helps control cell division, cell maturation, and cell death. Mutations (changes) in the gene that makes the MEK1 protein or in other genes in this signaling pathway may cause the MEK1 protein to become overactive in some types of cancer. This may increase the growth of cancer cells. Drugs that block the MEK1 protein are being used in the treatment of some cancers.




Principle

Differential Scanning Fluorimetry measures protein thermal unfolding by monitoring changes in fluorescence emission of a sample upon heating. This allows the determination of protein thermostability and complex formation even with weakly binding ligands by thermal shift assay. Differential Scanning Fluorimetry is therefore ideally suited for screening of optimum buffer conditions like pH, buffer composition and ionic strength. The technique is applicable to any biological sample, from soluble proteins to integral membrane proteins.

Applications

To identify low-molecular-weight ligands that bind and stabilize purified proteins.
To measure the denaturation and unfolding of proteins.

Procedure

1. Preparation of compound solutions
2. Preparation of buffer/additive screen plates
3. Preparation of compound storage plates
4. Equipment preparation
5. Sample preparation
26. Performing the scan

Materials

Real-time PCR instrument
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