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Analysis of Oligopeptidases B by Differential Scanning Fluorimetry(DSF) (CAT#: STEM-MB-0787-WXH)

Introduction

Oligopeptidases B (OpdBs) are two-domain serine peptidases with trypsin-like substrate specificity belonging to the prolyl oligopeptidase family. These enzymes are involved in the pathogenesis of trypanosomiasis, leishmaniasis, other parasitic infections, and in some cases bacterial infections and are potential targets for the development of therapeutic anti-infective agents.




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
41. Performing the scan

Materials

Real-time PCR instrument
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