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Analysis of Membrane Protein Stability by Differential Scanning Fluorimetry(DSF) (CAT#: STEM-MB-0781-WXH)

Introduction

Membrane proteins function in the cell as enzymes, receptors, and transporters as well as playing structural roles. With more than 50% of drugs on the market targeting membrane proteins, many are pharmacologically important. The 3D structure of a protein and the complexes it forms is invaluable in terms of informing how it works and for structure-based drug discovery.




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
35. Performing the scan

Materials

Real-time PCR instrument
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