I have been working with Santa Cruz antibody until for some reason labeling change in a bad way, so for that reason i decided try with another one, from ABCAM, after several trials with different timings and concentrations of primary antibody i haven't success,
In de data shed said that develops normal protocol, there is one picture anonymous and i already do the conditions and i am not able to replicate.
i would like to know if you have some advice, like incubation tempeture for first antibody, retrieval, special buffers? something?,
There is so much difference between incubate the primary ab at 4ºC that a room tenpeture?, is the retrieval important?, with one can i use? because i am working with cerebellum and the tissue it is extremely delicate more that forebrain.
I really appreciated any comments
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