A Discussion— If the polyclonal antibodies generated by antigen can be used to detect every subunits of the protein?


This is a common discussion on the full length protein. There are all kinds of opinions. If you are interested in this, you can join the discussion here and tell us your opinions on this topic.



The first speaker

In my opinion, I think if we immunize animals, such as, rabbit, with full-length protein as antigen, then the antigen generated in rabbit's serum is targeted at all aspects of this full-length protein. Especially the polyclonal antibodies, which can be targeted at the different subunits of protein. So I think the polyclonal antibodies generated during this process can be used to detect all kinds of subunit of the full-length protein. This is just a speculation. If it is right, when you are doing the experiment on detecting the subunits of full-length protein, you can take one kind polyclonal antibodies as the WB, and then you can tell the different subunits with the different molecular weight.

The second speaker

Technically, I agree with your opinion. However, there is a question that if the protein has three subunits, do the three subunits can generate the same quantity of antibodies? If not, the target of the pure protein will be wrong, which will lead to the uneven WB baseline, that is to say you can not get the detection results. What's more, we also can not determine the same condition of the different subunits.

The first speaker

For your opinion, the point is on the if you are comparing the different units. Then this will not be key in the balance of the different subunits.

The third speaker

Several years ago, I also was confused about this question. But finally I got it. Firstly, we should be clear with the definition on the protein. One protein is composed by several subunits. Actually, we always regard the single subunit as a protein, and the call the full-length protein as compound. Then we back to your question, as for the full-length protein, it is just a full length of protein which has a specific name, for the protein subunit, it is the full length of the subunit. As for the WB experiment, few people will study the whole subunits of proteins.

The first speaker

As for your(the third's) opinion, although we are talking about the full-length protein, when we are doing this, we always inject the quaternary structure of protein into the acceptors. So I think as for the full-length polyclonal antibodies, which contain the abilities of detecting the subunits. And when we are doing the experiment of SDS-PAGE, the only change is the dismissing of the subunits. The structure of the subunits does not change. So the polyclonal antibodies also have the ability to detect the different subunits.

The third speaker

In fact, there is no company will use the quaternary structure of protein to immunize animals. And in SDS-PAGE experiment, the subunit is separated.

These are the different opinions on the topic, how about yours? And if you have the different opinion, you can let us know.

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