Heat capacity studies of Zn-finger TFIIIA by Differential scanning calorimetry (DSC) (CAT#: STEM-MB-0485-WXH)

Introduction

Zinc finger (Znf) domains are relatively small protein motifs that contain multiple finger-like protrusions that make tandem contacts with their target molecule. Some of these domains bind zinc, but many do not, instead binding other metals such as iron, or no metal at all. Transcription factor IIIA (TFIIIA) is a very extensively studied eukaryotic gene specific factor. It is a special member of the zinc finger family of nucleic acid binding proteins with multiple functions.
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.

Add to Cart Please Inquire

Principle Applications Procedure Materials Notes Related Products

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

Other recommended products

Study of Lipid-protein interactions of hydrophobic proteins SP-B and SP-C in lung surfactant by Differential scanning calorimetry (DSC) (CAT#: STEM-MB-0525-WXH)
Study of Deuterium oxide stabilizing conformation of tubulin by Differential scanning calorimetry (DSC) (CAT#: STEM-MB-0511-WXH)
Studies of Thermal Denaturation of tRNAs by Differential scanning calorimetry (DSC) (CAT#: STEM-MB-0526-WXH)
Measurements of binding thermodynamics in drug discovery by Differential scanning calorimetry (DSC) (CAT#: STEM-MB-0522-WXH)
Self-Assembly Study of Supramolecular Nanostructures by Differential scanning calorimetry (DSC) (CAT#: STEM-MB-0503-WXH)
Isolatation and characterization of paracetamol form III by RHDSC (CAT#: STEM-MB-0506-WXH)
Detection of joint capsule changes by differential scanning calorimetry (DSC) (CAT#: STEM-MB-0510-WXH)
Analysis of Carbohydrates (Starch granules) by Differential scanning calorimetry (DSC) (CAT#: STEM-MB-0496-WXH)