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PBS in my Tris Glycine Running Buffer....what will happen


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#1 MKR

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Posted 14 January 2010 - 07:36 PM

So.... let's say an undergrad (not me of course) ran out of tris glycine sds running buffer and made more with the 10x PBS stock. So that about 1/7th of the running buffer was PBS. What do you think that would do???

#2 bob1

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Posted 14 January 2010 - 08:12 PM

So used PBS as the liquid component with added glycine, SDS etc? or just diluted PBS?

If just diluted PBS; then it won't work.

If with glycine, etc; it may work, but I suspect the ionic concentrations and odd buffering will do some weird stuff to the running conditions.

#3 mdfenko

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Posted 15 January 2010 - 08:20 AM

toss the buffer. the phosphate will affect the pH and the salt will cause problems when running the gel.

there is a gel formulation that uses phosphate buffer instead of tris-glycine (weber and osborn) but no salt, pH is 7 and uses the same buffer for the gel.

Edited by mdfenko, 15 January 2010 - 08:20 AM.

talent does what it can
genius does what it must
i do what i get paid to do

#4 MKR

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Posted 15 January 2010 - 10:26 AM

toss the buffer. the phosphate will affect the pH and the salt will cause problems when running the gel.

there is a gel formulation that uses phosphate buffer instead of tris-glycine (weber and osborn) but no salt, pH is 7 and uses the same buffer for the gel.


Yeah the buffer is tossed, but the samples were run for an hour before she realized what she had done (I'm not sure how this happened?). Anyway, it did run weird, but I'll go ahead with the western blot detection, as the marker seems generally alright (plus I can't start another one til Monday anyway).




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