We have decided to isolate the food vacuoles and examine the existence of proton pumps using a variety of antibodies.
exp.

Examine the results of this experiment.
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

The results below were obtained using primary antibodies against a common proton pump known to exist in paramecium (MW 56.1 kDa):


 

Legend
Lanes 1, 8  Size marker (kilo Daltons)
Lane  2      Purified proton pump
Lane  3      Lysate of the whole Paramecium
Lane  4      Lysate of the Parameciun food vacuole collected 0.5 min after feeding.
Lane  5      Lysate of the Parameciun food vacuole collected 5 min after feeding.
Lane  6      Lysate of the Parameciun food vacuole collected 10 min after feeding.
Lane  7      Lysate of the whole Paramecium reacted only with the secondary antibody.

Why did we use a sample of the purified proton pump (lane 2)?
Choose between right and wrong.

A. Using the purified protein we show that no other proteins can be recognized by our antibodies (no crossreactivity of the primary antibody with other proteins is detected).

B. The purified protein was used as a size reference that will indicate the size of the proton pump.

C. The purified protein serves to test whether the treated paramecia (exposed to yeast- containing medium for 30 sec) expresses proton pumps, as we previously hypothesized.

D. The purified protein is used as a positive control - it verifies the proper functioning of the antibodies and all other reagents.