Western blotting is a technique through which the sequences of amino-acid, present in the proteins can be identified. This method is pertinent to molecular biology. It also pertains to the cell. With the advancement of this technique, the biological researchers find it easy to recognize the amino-acids of complicated entangled proteins. To achieve the separation of particular amino-acids from a complex-structured protein, there are three necessary methods that need to be done. First of them is the separation by size. After performing the first step, the second step is to move the separated portion to a support, which is solid. The final part of the technique involves the marking of the selected amino-acids or proteins through the primary and secondary antibody. These antibodies are essential for the visualization (Liu, Mahmood, & Yang, 2014).
This technique is used by researchers for the separation and identification of proteins. If simply stated, the separation happens due to the molecular weights of proteins. Each protein has a fixed and distinct molecular weight. By identifying the masses of each protein in the cell, the separation becomes easy. The different portions or bands are formed for each type of protein. The outcomes we receive by separation through molecular weights are placed in the membrane. This membrane makes the section for every type of protein.
The way the amino-acids are recognized is the membrane, which has all the proteins according to their molecular weights, are separated from the ones that are desired by the researcher. The irrelevant antibodies are cleared and leave behind the ones that pertain the part of the research. The selected proteins are identified by the development of the film. Since these antibodies are separated in band form, the only band that is of the researcher’s interest remains. The size of the band tells the quantity of the present proteins.
Western blotting is a widely used technique to for the detection of the amino-acids or proteins. It also helps the researcher in the quantification of protein expression.
Liu, Z.-Q., Mahmood, T., & Yang, P.-C. (2014). Western blot: technique, theory, and troubleshooting. North American Journal of Medical Sciences, 6(3), 160.