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

Protein extraction from plant, animal, fungi, bacteria, etc is well known using different techniques. Many buffers are used to solubilize water-soluble proteins. There are few proteins which are insoluble and are rigid to be extracted by physical means or they lose activity when extracted by denaturants. One of the promising methods is to use organic solvents to extract proteins inactive state. C4 and C5 alcohols (butanol, pentanol) are mostly used for the purpose as they act as differentiating solvents. When an organic solvent is added to a protein solution followed by ammonium sulfate, the solution separates into three phases. Upper organic phase and lower aqueous phase, depending on the concentration of ammonium sulfate some proteins precipitate out and form an intermediate layer.

Immunocytochemistry is one of the most commonly used methods for protein localization in the cells or tissues using specific antibodies. The overall credibility of the results from Immunocytochemistry depends on two factors:

Mass spectrometric analysis is one of the important technique in protein biochemistry and peptide analysis. It is used to identify and analyze proteins and peptide samples. Care should be taken for sample preparation for mass spec analysis as the quality of data depends on the purity of sample applied. A variety of strategies are used to prepare the samples for mass spectrometric analysis.

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: