1. What is SDS page?
SDS PAGE also known as Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis is a technique used for separating the proteins based on their molecular weight. It is a widely used technique in forensics, genetics, biotechnology and molecular biology to separate the protein molecules based on their electrophoretic mobility.
2. What are the materials used in SDS page?
Power Supplies: It is used to convert the AC current to DC current.
Gels: These are either self prepared in the laboratory or are purchased from the market.
Electrophoresis Chambers: The chambers of the SDS-PAGE gels that fit well should be used.
Protein Samples: The protein is dissolved with SDS-PAGE sample buffer and boiled for 10 minutes. A reducing agent such as dithiothreitol or 2-mercaptoethanol is also added to minimize the disulfide linkages to prevent any tertiary protein folding.
Running Buffer: The protein samples filled on the gel are run in SDS-PAGE running buffer.
Staining and Destaining Buffer: Coomassie Stain Solution is used to stain. A destaining solution is used to destain a gel. Protein bands can then be observed with naked eyes.
Protein Ladder: A reference protein ladder is used to locate the protein of interest based on the molecular size.
3. What are the applications of SDS page?
The applications of SDS-PAGE are as follows:
It is used to measure the molecular weight of the molecules.
It is used to measure the size of the protein.
Used in peptide mapping
It is used to compare the polypeptide composition of different structures.
It is used to eradicate the pureness of the proteins.
It is used to separate the HIV proteins in HIV test
Analyzing the size and number of polypeptide subunits.
To analyze post-translational modifications.
4. What is the function of SDS in SDS page?
SDS can be defined as a detergent present in the SDS-PAGE sample buffer. The function of SDS is to break the disulphide bonds of proteins disrupting the tertiary structure of proteins along with some reducing agents.