Question

hello

how are you ?

I hope you are doing well

can you please help me to understand what is western bolting by writing a summary that describe a situation where you would need this technique and the steps of the procedure or how it works.

thank you so much

Answer 1

Western blotting was first described by Harry Towbin in 1979. It was in 1981 when W. Neal Burnette developed an improved version of the method and gave the name “western blotting” simply because of the location of laboratory.

Western blotting (protein blotting or immunoblotting) is a rapid and sensitive assay for detection and characterization of proteins. It is based on the principle of immunochromatography where proteins are separated into polyacrylamide gel according to their molecular weight.

Five steps are involved in western blotting and detection assay, namely, transfer, blocking, primary antibody incubation, secondary antibody incubation and protein detection and analysis. Proteins are moved from within the gel onto a membrane made of nitrocellulose(NC) or polyvinylidene difluoride(PVDF).

However, the Western blot is no longer used, and today the ELISA test is followed by an HIV differentiation assay to confirm HIV infection.

The western blot (sometimes called the protein immunoblot), or western blotting, is a widely used analytical technique in molecular biology, immunogenetics and other molecular biology disciplines to detect specific proteins in a sample of tissue homogenate or extract.