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.

While detergents play a vital role in separating proteins from the hydrophobic portions of the cell membrane during extraction and sample preparation, they need to be removed after successfully serving their purpose to prevent them from interfering with downstream applications (e.g., IEF, ELISA, protease digestion of proteins).

Successful biochemical analysis heavily relies on the effective extraction of biologically active proteins from source materials (e.g., cell and tissue samples). Thus, you need to have an excellent working knowledge of your target protein(s) and use the appropriate extraction buffers for a given experimental design to ensure optimal protein recovery.