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Monitoring RNA Stability by Differential Scanning Fluorimetry(DSF) (CAT#: STEM-MB-0762-WXH)

Introduction

Cellular RNA function is closely linked to RNA structure. The stability of an mRNA plays a major role in the determination of gene expression. The stability of an mRNA reflects its primary and higher-order structure, as well as its interactions with a variety of trans-acting RNA-binding proteins.




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
16. Performing the scan

Materials

Real-time PCR instrument