Polyacrylamide gel electrophoresis or SDS-PAGE is a widely used technique in molecular biology to separate and identify proteins. The technique involves denaturing the proteins with the use of Sodium Dodecyl Sulphate (SDS) and separating them based on their molecular weight.

In this blog, we will discuss the details of SDS-PAGE, its applications, and the role of Tri Sodium Sulphate in the process.

What is SDS-PAGE?

SDS-PAGE is a technique that involves separating proteins from a mixture based on their molecular weight through the use of Polyacrylamide gel. The proteins are first denatured with the help of SDS, a surfactant that binds to the protein and converts it to a uniform negative charge.

The denatured proteins are then loaded onto a Polyacrylamide gel, which separates the proteins based on their size. The smaller the protein, the quicker it will move through the gel via the electric field applied. By comparing the protein bands against a size marker, one can determine the size of the protein.

Applications of SDS-PAGE

SDS-PAGE is used in many applications of molecular biology. Here are a few:

- Protein Purification: SDS-PAGE aids in determining the purity of isolated proteins from cell lysates, chromatography columns, or any other source. The amount of protein can be measured by staining the gel with any protein-specific stain.

- Protein Identification: By using SDS-PAGE in combination with Western blotting, a protein's identity can be confirmed. This helps in detecting the presence of a particular protein in a sample.

- Protein Quantification: SDS-PAGE can also be used to quantify the amount of protein present in a sample. This can be done by comparing the protein bands against a standard protein to determine the unknown concentration.

Role of Tri Sodium Sulphate in SDS-PAGE

Tri Sodium Sulphate (TSS), also known as Sodium Tetraborate or Borax, is a critical component in the preparation of the running buffer required for SDS-PAGE. TSS helps in maintaining the pH of the buffer and stabilizing the electric field. Moreover, TSS inhibits the activity of proteases that can degrade the proteins during electrophoresis.

Conclusion

SDS-PAGE is an essential technique that every molecular biologist should know. It enables the separation of proteins based on their molecular weight, aiding in protein purification, identification, and quantification. Tri Sodium Sulphate plays a crucial role in maintaining the pH and stabilizing the electric field in SDS-PAGE. With this knowledge, scientists can isolate and study proteins from various sources, leading to significant discoveries in the field of molecular biology.