Western Blot Flashcards
1
Q
What biomolecules are detected in Western blot?
A
Proteins
2
Q
What is the mechanism of transfer of biomolecules to membranes in Western blot?
A
SDS coated negatively charged proteins are transferred from SDS-PAGE gel to membrane that is facing the positive electrode.
3
Q
What is the blocking reagent commonly used in Western blot?
A
Skim milk.
4
Q
What is the method to detect specific biomolecules in Western blot?
A
Primary antibody that binds to specific epitopes of the protein of interest. Secondary antibody that binds to constant region of primary antibody is conjugated to an enzyme eg. alkaline phosphatase. Light is produced at the vicinity of the secondary antibody when CSPD substrate is added.
5
Q
A
