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Heat capacity studies of SOX5 by Differential scanning calorimetry (DSC) (CAT#: STEM-MB-0484-WXH)

Introduction

Transcription factor SOX-5 is a protein that in humans is encoded by the SOX5 gene. This gene encodes a member of the SOX (SRY-related HMG-box) family of transcription factors involved in the regulation of embryonic development and in the determination of the cell fate. The encoded protein may act as a transcriptional regulator after forming a protein complex with other proteins. The encoded protein may play a role in chondrogenesis.
The energetics of formation of specific protein complexes with other proteins or nucleic acids is now attracting increasing attention since it actually represents a most basic biological function. The heat capacity plays a major role in the determination of the energetics of protein folding and molecular recognition. As such, a better understanding of this thermodynamic parameter and its structural origin will provide new insights for the development of better molecular design strategies.




Principle

Calorimetry is a primary technique for measuring the thermal properties of materials to establish a connection between temperature and specific physical properties of substances and is the only method for direct determination of the enthalpy associated with the process of interest. Calorimeters are used frequently in chemistry, biochemistry, cell biology, biotechnology, pharmacology, and recently, in nanoscience to measure thermodynamic properties of the biomolecules and nano-sized materials.
Amongst various types of calorimeters, differential scanning calorimeter (DSC) is a popular one. DSC is a thermal analysis apparatus measuring how physical properties of a sample change, along with temperature against time.1In other words, the device is a thermal analysis instrument that determines the temperature and heat flow associated with material transitions as a function of time and temperature. During a change in temperature, DSC measures a heat quantity, which is radiated or absorbed excessively by the sample on the basis of a temperature difference between the sample and the reference material.

Applications

Determination of equilibrium thermodynamic stability.
Study of folding mechanism.
Screening for thermal stability as an indicator for, ligand binding, pharmaceutical formulation or conditions conducive to crystal growth.
Study of the denaturation process.

Procedure

1. Instrument Start-up
2. Sample Preparation
(1) Dialyze the sample against the buffer that will be used as the reference for the experiment.
(2) Determine the concentration of the protein sample using the most suitable protein concentration determination method.
(3) Degas the sample and reference buffer in vacuum to get rid of microbubbles that can cause volume inaccuracy.
(4) load the samples and their respective buffer in pairs into 96 well plates compatible with the instrument.
(5) Place the plate in the sample holding compartment in the proper orientation.
3. Experimental Parameter Setup
Set the starting temperature, final temperature and the scan rate of the experiment.
4. Data Analysis

Materials

Differential Scanning Calorimeters
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