[Histonet] Tris Buffer Preservative

[Nicole Collette]

Hello, Amos,

It is pretty straightforward to make fresh, why would you want to 
store it? We use sterile 1M Tris, pH 7.4, and 4M NaCl as stocks, they 
are concentrated enough that nothing obvious grows in them, dilute 
the volume you need, pH if needed, (add the Tween if needed), and 
store in the fridge for a single experiment. Alternatively, you can 
make a sterile 10X salt stock, dilute that as needed to make the math 
easier. Throw out the leftover diluted solution so nobody else tries 
to use it. In my experience, TBS(T) will ALWAYS get cloudy when left 
to sit, I wouldn't risk the contamination. However, I'm not in a 
high-throughput lab, so this works for me (I'm a one-pair-of-hands 
histology lab).

Sincerely,
Nicole Collette
Lawrence Livermore National Lab/ UC Berkeley


At 11:59 AM -0500 2/24/10, Amos Brooks wrote:
>Hi,
>    I have been making up my own tris buffer for immunohistochemistry. It has
>been working swimmingly, with one exception. I have been doing some work on
>Salmonella, and apparently the researcher has been seeing bacteria in her
>immunofluorescent slides that she says are moving and seem to be viable. She
>wanted to culture all my reagents. So the primary and secondary antibody
>diluents (commercially purchased) came up clean, but the TBS ended up with a
>pretty healthy growing colony. Now since I don't do IHC overnight in an
>incubator, I don't think this is necessarily a catastrophy. (No one else has
>noticed this) It does seem to warrant further investigation though. So for
>the folks that make their own solutions up, what do you use as a
>preservative for your buffers and how much do you use. I haven't seen
>anything in any of the recipes I have found. I was thinking Sodium Azide,
>but it is really hazardous, and Wikipedia says it is actually explosive (
>http://*en.wikipedia.org/wiki/Sodium_azide). Biocare Medical's data sheet
>says they use less than 0.25% procyclin. (thanks for not hiding the
>ingredients Biocare, I love you for that!) Has anyone tried that? Any other
>suggestions would be welcome.
>
>Thanks,
>Amos
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