Macromolecule Blotting and Probing

Questions and Answers

To isolate DNA and RNA

To identify proteins and nucleic acids (correct)

To synthesize proteins and nucleic acids

To visualize gel electrophoresis

Southern blotting

Western blotting

Eastern blotting

Northern blotting (correct)

To visualize protein detection

To transfer proteins from gel to membrane (correct)

To detect proteins using antibodies

To separate proteins by size

To prevent non-specific binding Signup and view all the answers

Central blotting Signup and view all the answers

To detect proteins using antibodies Signup and view all the answers

By using mechanical innovation, such as homogenizer Signup and view all the answers

To prevent the digestion of the sample at cold temperatures Signup and view all the answers

Study Notes

Blotting is a method that transfers nucleic acids (RNA and DNA) or proteins onto a membrane, which can be made of nitrocellulose, polyvinylidene difluoride (PVDF), or nylon.
The membrane is used to visualize the transferred molecules using staining, with examples including Ethidium bromide, Crystal violet, Safranine, and Ossmium tetroxide.
Blotting is used in molecular biology to identify proteins and nucleic acids and is widely used for diagnostic purposes.

Principle: Western Blotting isolates proteins by size and specifically identifies proteins of interest using a counteracting agent.
The procedure involves six steps: sample preparation, gel electrophoresis, protein transfer, blocking, antibody incubation, and protein detection and visualization.
Western Blotting is a widely accepted scientific method used to identify specific proteins.

Proteins can be extracted from samples, such as tissues or cells.
Tissue samples are broken down using mechanical innovation, such as homogenizers, and protease and phosphatase inhibitors are used to prevent digestion at cold temperatures.
Protein concentration is detected using a spectrophotometer, which allows for the mass of proteins loaded into each well.