how to color the stacking gel?

[Iraz Toprak Aydin]

Hello!

I have seen some SDS PAGE gels with red or orange stacking part that makes
the wells visible. They use phenol red for the red stacking gel. But I
couldn't find the concentration of phenol red they use for this purpose.
Does anybody have any experience in this?

Thanks in advance!

Iraz Toprak Aydin

EPFL SV ISREC, Station 19

Batiment SV, SV 2540

CH-1015 Lausanne

Switzerland

Tel: +41 21 693 07 36

e-mail: <mailto:iraz....@epfl.ch> iraz....@epfl.ch

[Ben Thuronyi]

This page recommends 0.012% bromophenol blue in the upper stacking gel for this purpose, and has photos.

http://bitesizebio.com/2010/03/22/sds-page-the-easy-way-to-find-the-wells/

~ Ben T.

Hello!

Thanks in advance!

Iraz Toprak Aydin

Batiment SV, SV 2540

CH-1015 Lausanne

Switzerland

e-mail: <mailto:iraz....@epfl.ch> iraz....@epfl.ch

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[WS]

Hi,

just add as much as you like. I have saturated solution of phenol red
(some material not dissolved) in water, shake it well before use and
the add 1 or 2 drops. You may also add it to the separation gel, as
you then may monitor the front very easily. In contrast to
bromophenolblue phenol red polymerizes to some extent into the gel,
then it's easier to locate and remove air bubbles in blots.

Have fun!

Wo

[Dr Engelbert Buxbaum]

In article <mailman.660.127419...@net.bio.net>,
iraz....@epfl.ch says...

>
> Hello!
>
>
>
> I have seen some SDS PAGE gels with red or orange stacking part that makes
> the wells visible. They use phenol red for the red stacking gel. But I
> couldn't find the concentration of phenol red they use for this purpose.
> Does anybody have any experience in this?

See

@ARTICLE{Smi-88,
author = {I. Smith and R. Cromie and K. Stainsby},
title = {Seeing gel wells well},
journal = {Anal. Biochem.},
volume = {169},
year = {1988},
pages = {370-371},
doi = {10.1016/0003-2697(88)90297-7},
language = {engl},
}

Note however, that this works only with negative detergents like SDS. If
you do electrophoresis with positively charged detergents like CTAB, you
need a different dye:

@INPROCEEDINGS{Bux-09,
author = {E. Buxbaum},
title = {Cationic electrophoresis and eastern blotting},
chapter = {14},
pages = {115-128},
editor = {B.T. Kurien and R.H. Scofield},
booktitle = {Protein blotting and detection -- {M}ethods and
protocols},
publisher = {Humana},
year = {2009},
volume = {536},
series = {Meth. Mol. Biol.},
isbn = {978-1-934115-73-2},
address = {Dordrecht},
language = {engl},
}