Buffer components, Blotting buffers – Bio-Rad Mini-PROTEAN® TGX™ Precast Gels for 2-D Electrophoresis User Manual
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TBE-urea (30 ml)
89 mM Tris, 89 mM boric acid, 2 mM EDTA, pH 8 .0, 12%
(catalog #161-0768)
Ficoll, 0 .01% bromophenol blue, 0 .02% xylene cyanole,
Store at 4°C
7 M urea
Tris base
0.32 g
Boric acid
0.165 g
EDTA
17.5 mg
Ficoll
3.6 g
Bromophenol blue
3 mg
Xylene cyanole FF
6 mg
Urea
12.6 g
diH
2
O
to 30 ml
Buffer Components
0 .5 M Tris-HCl, pH 6 .8 (1 L)
Tris base
60.6 g
(catalog #161-0799)
diH
2
O
~900 ml
Store at 4°C
Adjust to pH 6.8 with HCl
diH
2
O
to 1 L
10% SDS (250 ml)
SDS
25.0 g
(catalog #161-0416)
diH
2
O
to 250 ml
1 .0% Bromophenol blue (10 ml)
Bromophenol blue
100 mg
(10 g powder, catalog #161-0404)
diH
2
O
to 10 ml
Blotting Buffers
Towbin buffer (1 L)
25 mM Tris, 192 mM glycine, 20% methanol
Dissolve:
Tris base
3.03 g
Glycine
14.4 g
diH
2
O
500 ml
Then add:
Methanol
200 ml
diH
2
O
to 1 L
Alternatively, use:
10x Tris/glycine (catalog #161-0734)
100 ml
Add 200 ml methanol and diH
2
O to 1 L as above
Tris-buffered saline with
20 mM Tris, 500 mM NaCl, 0 .05% Tween 20
Tween (TTBS, 1 L)
Tris base
2.4 g
NaCl
29.2 g
10% Tween 20
5.0 ml
diH
2
O
to 1 L
Alternatively, use:
10x TBS (catalog #170-6435)
100 ml
10% Tween 20 (catalog #166-2404)
5 ml
diH
2
O
895
ml
Instruction Manual and Application Guide