testing antibody dilution
I'm trying to figure out the right dilution for my antibody stained in human
liver cell-line. I ran several dilution and along with a positive control.
I'm expecting my antibody to be located in nucleus. Positive control showed
it's so and it also stained in cytoplasmid. On my human liver cell-line it
showed no nucleus stain (good for me). However, there is very fainted
stained on the cytoplasmid. Two dilution of primary antibody I don't know
what I can conclude as my good dilution for my antibody when compare with
the positive control.
1) I antibody at 1:5000 showed on a + control showed a complete staining in
nucleus and in cytoplasmid. However, negative human liver cell-line showed a
no nucleus staining (good for us), but very fainted here and there in the
2) I antibody at 1:7000 showed on a + control showed a complete staining in
nucleus (but not strong) and in cytoplasmid. However, negative human liver
cell-line showed a no nucleus staining (good for us), as well as in the
My question is, what would be my recommendation dilution for my antibody
that I have tested after compared with my positive control. Should I go for
Any opinion or advice would be more appreciated.
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