I just realized that I incubated my sections with two primary antibodies both of which are raised in mouse. I don't want these sections to go to waste and am hoping to wash away any bound primary antibodies so I can apply the correct primaries (one raised in mouse, the other in rabbit).
Is there any way of doing this?
Note: These primaries are not conjugated and so require application of respective secondary antibodies.
As part of multiplex staining protocols, we do what is called 'stripping' and removing previously bound primary Ab.
There are a few solutions you can use (ref: Article Antibody Elution Method for Multiple Immunohistochemistry on...
)but a shortcut is to run your low pH HEIR step again, as we already have that validated.
I believe you can follow this protocol Ranjan 2012, doi:10.1016/j.jim.2012.02.018. It is Tris buffer, pH 6.8 with 2% SDS and 20 mM DTT, incubate a few hours at RT .
There is another protocol explained in
Micheva KD, Busse B, Weiler NC, O'Rourke N, Smith SJ.
Neuron. 2010 Nov 18;68(4):639-53. doi: 10.1016/j.neuron.2010.09.024.
PMID: 21092855
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