Dear Histonet,

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 
cytoplasmid.
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 
cytoplasmid.

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 
the 1:5000?

Any opinion or advice would be more appreciated.

Thao

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