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)

Introduction

From the time an infant takes his first breath, pulmonary surfactant forms a continuous film at the liquid-air interface of the alveoli, which is fundamental to breathing. This film, through its surface-tension lowering properties, greatly decreases the work of breathing and imparts a remarkable stability to the alveolar cavities, thus preventing their collapse under the physiological duress of the breathing cycle. The surfactant proteins B (SP-B) and C (SP-C) are important for the stability and function of the alveolar surfactant film.

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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

Study of Lipid-protein interactions of hydrophobic proteins SP-B and SP-C in lung surfactant.

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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