Question

Questions refer to Western Blots

2. MLC1 is approximately 22kD, myosin heavy chain is 200 kD, and actin is 42kD. Which protein will migrate fastest through the gel, why?

3. Why are proteins blotted from the gel onto the membrane?

4. State two pieces of information the western blot result provides about the protein of interest.

Answer 1

2. Lightest one would travel fastest. Hence MLC1 would travel farthest, followed by actin and myosin.

3. Proteins are blotted on the membrane for the detection of specific protein via specific antibodies. We can not hybridize antibodies with protein on PAGE gel because it has pores and bands can diffuse over time hence we will get smears instead of clear bands. Again the gel has come in contact with various proteins hence there can be non-specific binding of antibodies.

4. We can identify selective protein among a mixture of proteins by using specific antibodies against the protein. We can measure protein expression and relative amount of protein expression.