Bio-Rad Trans-Blot® Plus Cell - Manual (Page 22)

Dunn carbonate buffer
In some cases, this buffer may produce higher efficiency transfers and improve
the ability of antibodies to recognize and bind to proteins.
10 mM NaHCO
3
, 3 mM Na
2
CO
3
, 20% methanol, pH 9.9
a) Using dry reagents:
0.84 g NaHCO
3
0.318 g Na
2
CO
3
(anhydrous)
500 ml deionized water (dd H
2
O)
200 ml methanol
add dd H
2
O to I liter
2. Buffers for native gels
Towbin Buffer
25 mM Tris, 192 mM glycine, pH 8.3
a) Using 10X Tris /glycine buffer (catalog #161-0734 for 1L bottles or
catalog # 161-0757 for 5L cube)
100 ml of 10X Tris /glycine buffer
900 ml of deionized water (dd H
2
O)
b) Using dry reagents:
3.03 g Tris
14.4 g glycine
600 ml deionized water (dd H
2
O)
add dd H
2
O to 1 liter
3. Buffers for nucleic acid transfers
TBE (Tris-Borate EDTA)
89 mM Tris borate, 2 mM EDTA pH 8.3
a) Using 10X TBE buffer (catalog #161-0733 for 1L bottles or
catalog # 161-0770 for 5L cube)
100 ml of 10X TBE buffer
900 ml of deionized water (dd H
2
O)
b) Using dry reagents (10X stock)
108 g Tris base
55 g boric acid
40 ml 0.5 M EDTA, pH 8.0
Add 100 ml of the 10X stock to 900 ml deionized water (dd H
2
O)
to make a 1X working solution.
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