2 replies to this topic

[Praitibha Batra]

Hi everyone...I am looking for an alternative chemical to be used for beta alanine in the acidic native PAGE experiment. Could anyone please enlighten me on this topic?And what is the subtle difference between beta alanine and L-alanine could I use the latterr coz that one is available.And an alternative for malachite green loading dye fr acidic native PAGE,I do not have that one also.Thank you in advance.

[Praitibha Batra]

Hi I also need the appropriate resolving and stacking gel compositions for different percents 12% and 15%.thank you.

[mdfenko]

beta-alanine is a beta amino acid, l-alanine is an alpha amino acid (position of amine relative to carboxylic acid group). you can look at the wikipedia page about beta-alanine.

to replace beta-alanine in the running buffer, you must find a buffer with a similar pK that won't otherwise react with the samples. malic acid may fit the bill, but you should check buffer tables.

as for tracking dye, you could use bromphenol blue if you don't mind looking for a yellow band (i think it actually remains blue at the running pH of 4.3).

the stacking gel formulation (2.5% acrylamide, 0.625% bis-acrylamide) should not be adjusted for different percentage separating gels.

the recipe i have (and think have posted someplace on bioforum) makes a 15% separating gel. just adjust the acrylamide and water to make 12% (reduce acrylamide volume by 1/5th and increase water by the difference in acrylamide volume)

Edited by mdfenko, 08 March 2013 - 05:21 AM.