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how to dilute monoclonal antibody?

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Dierk:
Hi,

I have a monoclonal anti-tyrosine hydroxylase AB (Sigma T2928)

Now I wonder how I should dilute this antibody:

in aqua dest (pH-adjusted or not)?
in tris?
in PBS?

I heard some people say that sometimes tris doesn't work well...

What about dilution factor? 1:10? 1:100? 1:250?

I would dilute the AB and store it all at -25 °C

so some people say to much of a dilution is no good because the protein will be "too lonely" in ice water and therefore break down more rapidly...


so what do you suggest?

We want to use this AB for 1 or 2 years at least.

thx,
Dierk

richard03:
Dierk,

First, you should read the Ab datasheet and follow instruction on that sheet. PBS is commonly used buffer for diluting Abs if you cann't find the info on datasheet.

If you want to store your Abs for long term, I recommend to aliquat (at least 10ul each) and store at concentrated form. This is the best way to achieve long term storage of antibodies.

However, some antibody's working dilution is pretty high such as TH (1:1000~5000). I don't know for the one from Sigma you currently have, but Chemicon's TH antibody usually 1:2000 and up. In this case you can dilute the antibody 1:10 and no further. Then aliquat and store at -80C that should be fine for TH and most of other antibodies.

One exception I have encountered was Dopamine D1 receptor Ab, that is not  good after storage at -80C for a few months. So try to read info from datasheet that usually tells you what is the best storage temp.

Richard

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