Pre-prac DAY 2 - Western Blot interactive

Question 1

What other lab method is commonly used before a Western Blot? Why?

Answer 1

SDS-PAGE to separate proteins by weight via electrophoresis

Question 2

What ‘force’ is used to transfer the proteins from an SDS-PAGE run onto a porous membrane for use in Western blotting?

Answer 2

an electric field (i.e. electricity)

Question 3

What other name is commonly given to Western blotting?

Answer 3

Immunoblotting (and protein blotting)

Question 4

What is the role of primary antibodies (w.r.t. Western blotting)?

Answer 4

Specific binding to the protein of interest which can then bind to the membrane

Question 5

What is the role of secondary antibodies (w.r.t. Western blotting)?

Answer 5

Binds specifically to the primary antibody and is bound to an enzyme which can be used to detect the protein of interest