Analysis of Ligands of Thermophilic ABC Transporters by Differential Scanning Fluorimetry(DSF) (CAT#: STEM-MB-0784-WXH)

Name: Analysis of Ligands of Thermophilic ABC Transporters by Differential Scanning Fluorimetry(DSF)
SKU: STEM-MB-0784-WXH
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Introduction

ATP-binding cassette (ABC) transporters are a large superfamily of membrane proteins with diverse functions. They convert the energy gained from ATP hydrolysis into trans-bilayer movement of substrates either into the cytoplasm (import) or out of the cytoplasm (export).

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Principle Applications Procedure Materials

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
38. Performing the scan