1. explain the principal behind western blotting and Elisa ? write the difference between Native and...

1. explain the principal behind western blotting and Elisa ?

write the difference between Native and the SDS PAGE?

Expert Solution

1. Principal of western blotting

Western blotting is used to identify specific proteins that is present in the given sample. It is also called as immunoblotting because it used to identify antigen. Western blotting is based on two principles: SDS-polyacrylamide gel electrophoresis and protein blotting. SDS electrophoresis is based on the principle that biological charged particles moved under the influence of electric fields. Here, proteins are separated based molecular mass. In this electrophoresis, SDS which is a detergent is used

After separation, separated proteins are blotted onto a matrix which is made up of nitrocellulose membrane and stained with antibodies that act as a probe to specific target proteins.

Principal of ELISA

ELISAs or (Enzyme-linked Immunosorbent Assays) based in the principle that specificity of antibodies with the simple enzyme assays sensitivity, with the help of antibodies or antigens which is coupled to an easily-assayed enzyme.

Difference between Native and the SDS PAGE is:

In Native PAGE separation of biomolecules are based on charge, size and shape. This page is useful for purification and separation from protein mixture.

In SDS PAGE separation of biomolecules is based on the only on molecular weight of proteins. It is to measure purity of proteins.

gladiator answered 2 years ago

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