Question

1. How does immunoblotting allow for a specific protein to be identified?

2. The primary antibody was made by injecting purified Rubisco proteins into rabbits and collecting the antibodies that the rabbit produced against this foreign protein. The secondary antibody was made in a goat. What was injected into the goat?

3. What would have happened if the membrane was not incubated in blocking buffer before the primary antibody incubation step?

4. During the transfer step, what would have happened if the gel and membrane were reversed in the order of assembly?

Can you answer this questions in details and explain it please

Answer 1

A specific protein is recognized by specific antibody which can be tagged with a enzyme and then if you add substrate it will give colour.the antibody can be fluorescence tagged also.

2)the primary antibody was injected into the goat.which is give anti rabbit secondary antibody.

3)the all the membrane can be bound by the primary antibody and thus a defected result will come. Nonspecific binding will be seen..

4)if you reversed the membrane and gel the. Protein will move freely to outside of the gel into the solution because protein will be transferred to positive side .

And membrane will not get any protein molecules.