The Protein Man's Blog | A Discussion of Protein Research

Protein samples are analyzed by denaturing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Total protein lysates or protein samples from purification experiments are separated on 1D and 2D gels depending on their molecular weights and PI.  Appropriate staining techniques are used to detect proteins on SDS-PAGE gels depending on the protein of interest, downstream experiments like Western blotting, mass spec analysis etc.,

While researchers use several ways to visualize their protein samples, there isn’t a single best approach that is ideal for all cases. Each visualization technique has its own unique advantages and limitations, and so may only be suitable for certain applications.

Proteomics is a complex field and it is exposed to major developments over the last few decades. A general proteomic experiment is a multi-step process, which involves modifications and analysis at every step depending on the experimental requirement. Crudely, these experiments have the following steps:

While several protein separation technologies have been developed in recent years, two-dimensional gel electrophoresis remains one of the most popular techniques used in the field of proteomics. This is not surprising since 2D PAGE is routinely used to accurately analyze the individual components of even the most complex proteins, and can simultaneously resolve more than 5000 proteins (depending on the gel size used).