Question

What should your coomassie result tell you about your taq prep that you cannot learn from the western blot? (Biotechnology)

Answer 1

Them two are staining strategies:

Coomassie Brilliant Blue for General protein gel stains

A few colors have been presented for protein gel coloring. These mixes tie firmly to proteins and ingest light in the obvious area of the range. There is no single completely best compound. The decision of the color depends for the most part on the amount of the protein to be identified. The accompanying colors are recorded in the request of expanding affectability: Coomassie Brilliant Blue R 250, Acidic Fast-Green, Amido dark, and silver nitrate.

Coomassie Brilliant Blue is the most often utilized protein gel coloring color. Contingent upon the thickness of the gel and the properties of the protein, as low as 0.1 microgram protein can be distinguished utilizing this color.

Particular protein identification techniques: Western blot

In the event that, rather than distinguishing all proteins, the objective is the very particular discovery of a specific protein, two unique strategies are accessible.

One strategy called Western blot can be utilized if an exceedingly specific counter acting agent is accessible against the protein of intrigue. The guideline of the method is as per the following. In the initial step, proteins are isolated in view of their size by utilizing SDS-PAGE. Along these lines, a nitrocellulose film is laid firmly on one side of the gel. This "sandwich" is put into an extraordinary electrophoresis mechanical assembly. By performing electrophoresis toward a path opposite to the plane of the gel, the proteins will be exchanged from the gel onto the film. This progression protects the relative spatial course of action of individual protein groups.

The film conveying the firmly bound proteins is then hatched in an answer containing the particular counter acting agent against the objective protein, for the most part called the essential immune response. The essential immune response will tie just to the objective protein. To imagine the destinations where the immunizer has bound to the layer, an optional counter acting agent is included that had been conjugated (covalently bound) to a compound. The optional immunizer perceives the consistent locale of the essential counter acting agent. Consequently, a given optional counter acting agent conjugate can be utilized with numerous essential antibodies.

After this progression, the film is absorbed an answer containing a substrate of the conjugated compound. Reactant transformation of the substrate yields a beautiful insoluble item that encourages at the site where it was created and, at last, marks the site of the objective protein.