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Analysis of Ricin Inhibitors by Differential Scanning Fluorimetry(DSF) (CAT#: STEM-MB-0825-WXH)

Introduction

Ricin is a natural, highly toxic compound that is a byproduct of processing castor beans. If inhaled, injected or ingested, less than a pinpoint of ricin can kill a person within 36 to 48 hours due to the failure of the respiratory and circulatory systems. Ricin is a potent cytotoxin easily purified in large quantities. It presents a significant public health concern due to its potential use as a bioterrorism agent. For this reason, extensive efforts have been underway to develop antidotes against this deadly poison.




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
79. Performing the scan

Materials

Real-time PCR instrument
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