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Analysis of HIV-1 Capsid Protein by Differential Scanning Fluorimetry(DSF) (CAT#: STEM-MB-0786-WXH)

Introduction

Functional core structure consisting of numerous capsid (CA) proteins is a major component of viral particles and is essential for the replication of human immunodeficiency virus type 1 (HIV-1). As well-documented, Gag-CA plays critical multiple roles at various steps in the HIV-1 life cycle. It needs to be underscored that the biological and/or biochemical analysis from different angles of this multi-functional viral protein is a prerequisite to understand the virology of HIV-1 with a complicated replication mode.




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
40. Performing the scan

Materials

Real-time PCR instrument
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