Western blotting is a key technique in cell and molecular biological research. Researchers can identify specific proteins in complex proteins by using western blot. You can know more about western blot service online via https://www.bosterbio.com/services/assay-services/western-blotting-service.
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To accomplish this task, the technique relies on three components:
(1) separation by size
(2) transfer to a solid substrate
(3) marking target proteins using proper primary and second antibodies to visualize.
This paper will explain western blot and provide some solutions. Research often uses Western blot to identify and separate proteins. This technique separates proteins based on their molecular weight and then by type using gel electrophoresis.
The results of this procedure are transferred to a membrane that creates a band for each individual protein. After the membrane has been incubated with antibodies specific for each protein, it is removed from the incubator.
The unbound antibody is removed and only the bound antibody to your protein of interest is left. The film is then developed to detect the bound antibodies. Only one band should be visible as the antibodies only bind the protein of particular interest.
The amount of protein in the band determines its thickness. Thus, a standard can be used to indicate how much protein is present. This paper will describe the western blot protocol.
It will also include pictures and theories to aid in understanding the protocol. The procedure will then be explained theoretically, with the final section containing troubleshooting tips.