If I were to use an primary antibody for immunohistochemistry for a specific protein, the secondary antibody should be the anti-host of the primary antibody.
For example, if I am using a human/mouse/rabbit antibody with sheep as a host as a primary antibody. The secondary needs to be anti-Sheep. But if my primary antibody is polyclonal, can I use anti-mouse/rat/donkey/rabbit/goat as secondary antibody ?
OR
Do i have to see the homology % for the protein between the host of primary and secondary for the usage?
Antibody are termed as monoclonal or polyclonal due to specific epitope-paratope interaction which is decided by fab region of primary antibody. Since secondary antibody binds to fc region of primary antibody, which is species specific. Hence a anti sheep secondary antibody should be preffered. If this particular type is not available and, you are using secondary antibody raised in another species, always run a negative control( in which primary antibody is ommited) to see how specific your secondary antibody works.
Hope this might help.
You must use anti pry ab+ as secondary ab. If it is either monoclonal or polyclonal.
You have to use an anti-host secondary antibody. For example, if you are using a human or mouse primary antibody, which is raised in sheep, then the secondary should be anti-sheep antibody only.
One more example, if you have a mouse sample, use primary anti-mouse (raised in rabbit) , then the secondary will be an anti-rabbit secondary antibody (secondary can be raised in sheep or goat or donkey anything doesn't matters).
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