Here is my usual protocol for native page:
Separating gel buffer contained 8% acrylamide (aa:bis30%:0.8%) in 1.5 M acetate-KOH (pH 4.5), plus 70 μl 10% ammonium persulfate (APS) and 2.5 μl N,N,N,N′-tetramethylethylenediamine (TEMED) per gel (5 ml) to start polymerization.
The stacking gel contained 3% acrylamide, 0.25 M acetate-KOH (pH 6.5), 20% glycerol, and 30 μl 10%APSand5μl TEMED per gel (2.5 ml) to start polymerization.
The running buffer contained 0.35 M β-alanine, and 0.14 M acetic acid(pH 4.5).
I don't have beta-alanine right now. Can I substitute it with 6 amino hexanoic acid?
I used for running buffer 50mM Bistris/HCl pH 7.0 and 750 mM 6 amino hexanoic acid.
Thanks Elvira,
I precise that I am not talking about Bleu native PAGE but acidic native gel for "basic" proteins with pI above 7. Was your answer concerning BN PAGE or Native acidic (or basic) gels ?
Why don't you search about pKa of Beta alanine(3-amino propanoic acid?) and 6-amino hexanoic acid? How about glycine?
6-amino hexanoic acid pK1 4.43; pK2 10.75
3-amino propanoic acid pK1 3.6; pK2 10.9
Running buffer being at pH 4.5 might not make a big difference
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