Analysis of Egg Yolk Proteins and Their Thermal Sensitivity by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) (CAT#: STEM-ET-0337-ZJF)

Introduction

The multiple functional properties of egg yolk are mostly influenced by its complex protein composition. The high lipid content of egg yolk as well as the low solubility of delipidated egg yolk lipoproteins make analysis by conventional chromatographic or electrophoretic techniques a difficult task. This service provides an SDS polyacrylamide gel electrophoresis (SDS-PAGE) method to profile egg yolk proteins after delipidation with acetone using sodium dodecyl sulfate polyacrylamide gel electrophoresis on precast 8−18% T polyacrylamide gradient gels. The bands are identified based on their molecular weight and by comparison with isolated egg yolk subfractions. This method is very well suited for assaying the thermal sensitivity of whole yolk and its components and thus for the characterization of heat treatment processes.

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Principle Applications Procedure Materials Notes Related Products

Principle

Polyacrylamide gel electrophoresis (PAGE) is an electrophoresis which based on polyacrylamide gel. Polyacrylamide gel is tougher than agarose gel and is relatively inert, thermostable, strong, and transparent, with a uniform pore size. PAGE is a widely used technique for separating and analyzing proteins based on their size and charge. It involves the migration of proteins through a porous gel matrix made of cross-linked polyacrylamide under the influence of an electrical field. Known as one of the most versatile and widely used techniques, PAGE has the ability to separate proteins at high resolution, allowing the detection of minor differences in protein composition.
In sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), sodium dodecyl sulfate (SDS) is added to PAGE. SDS-PAGE eliminates the influence of structure and charge in electrophoresis, and proteins are separated based on the differences in their molecular weight.

Applications

Agricultural and food chemistry, proteins, thermal denaturation

Procedure

1. Preparation: Prepare electrophoresis buffer with a certain pH. Prepare a gel solution of appropriate concentration, taking care not to produce bubbles, and use a cast to solidify the solution into a gel.
2. Sample Application: Put the prepared gel with a cast into the electrophoresis tank, and add an appropriate amount of buffer. Pipette the sample into the sample wells.
3. Electrophoresis: Adjust the appropriate distance between the electrodes. Switch on the electrophoresis apparatus and set the voltage and current. Perform electrophoresis for a period of time.
4. Determination: After electrophoresis, stain the gel for visible results. Observe, record and analyze the position of separated bands. Or, utilize an imaging system for analysis.
5. Downstream processing: After separation, an additional method is often applied to the separated bands for further processing. For example, cut the band out of the gel as a slice to dissolve and purify it.

Materials

• Gel electrophoresis apparatus
• Sample solution
• Buffer solution and SDS-polyacrylamide gel

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