The protein on the membrane can either be detected by the use of a reporter-labeled primary antibody directed against the protein or a reporter-labeled secondary antibody directed at the primary antibody.

The transfer of the protein is known as blotting.

The proteins are then transferred to a nitrocellulose or polyvinylidene fluoride (PVDF) membrane, where they are immobilized.

The proteins used for western blotting are separated by gel electrophoresis to obtain them on a gel matrix.

The principle of western blotting is the interaction between the proteins and the probes used for the detection of the proteins.

Western blotting has a number of advantages over other similar techniques as the process only requires the use of a small amount of reagents, and the same protein transfer can be used for multiple analyses.

Unlike Southern blotting that utilizes radio-labeled nucleic acid probes, western blotting usually uses a second antibody tagged with an enzyme.

Western blotting or protein blotting depends on the specificity of interaction between the protein of interest and the probe used for the detection of the protein.

The sensitivity of the process depends on the efficiency of transfer retention of proteins during processing and the final detection.The immunodetection of proteins has a wide application in biochemistry and other sciences as it can detect and characterize a multitude of proteins.

Western blotting has revolutionized the field of immunology with the use of antibody probes against membrane-bound proteins.The proteins can then be identified on the membrane by different means. The process of western blotting involves the transfer of proteins separated by SDS PAGE into an absorbent membrane.

The use of polyacrylamide gel electrophoresis is a prerequisite for western blotting in order to separate proteins prior to their identification.Western blot, also known as immunoblotting, is the process of separating proteins and identifying them in a complex biological sample.

Western blot vs southern blot