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Analysis of Lectins by Differential Scanning Fluorimetry(DSF) (CAT#: STEM-MB-0804-WXH)

Introduction

Lectins are defined as proteins that bind to carbohydrates. The same features that lectins use to defend plants in nature may cause problems during human digestion. They resist being broken down in the gut and are stable in acidic environments, features that protect lectin-containing plants in nature.




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
58. Performing the scan

Materials

Real-time PCR instrument