C.B.S. Scientific EBU-102P - Manual (Page 12)

C.B.S. œ Scientific
12
Electro-Blotting System
SECTION 4
Applications & Running Conditions for Tank Type Electro-Blotting
4.1
Recommended Buffers, Membranes, Power Settings and Transfer Times
Gel Type
Blotting Buffer Transfer Membrane Power
denaturing:
SDS-PAGE
(Western) Towbin (25mM Tris/192mM Glycine/
pH 8.3/Methanol 20% (w/v) nitrocellulose/nylon
PVDF: ion-exchange •
room temperature, 70V-
100V, 200mA, 1.5- 4
hours
•
4ºC, 150V, 400mA, .5-2
hours
•
Overnight, 30V, 75mA
non-denaturing:
SDS-PAGE
(acidic or neutral
proteins) 25mMTris/192mM Glycine, pH 8.3
25mM sodium phosphate, pH6.5
15mM sodium borate, pH9.2 nitrocellulose; nylon
PVDF; ion-exchange
diazo-paper
diazo-paper •
room temperature, 60V,
200mA, 4 hours
•
4ºC, 125V, 400mA, 1.5
hours
•
Overnight, 30V, 100mA
IEF (native-
basic proteins,
acid urea gels) 0.7% acetic acid (w/v)* nitrocellulose; nylon
diazo-paper
•
room temperature, 70V,
400mA, 4 hours
•
4ºC, 100V, 600mA, 1.5
hours
•
Overnight, 30V, 200mA
DNA
(Southern) 1XTAE or agarose
0.5X TBE, acrylamide nylon; +charged
nylon, neutral or
+charged •
room temperature, 30-
40V, 25-150mA, 2-4
hours
•
4ºC, 125V, 400mA, 1.5
hours
RNA
(Northern) 19.6mM phosphate/5.4mM
citrate
pH3.0 or 25mM sodium
phosphate
pH 6.5 nylon nitrocellulose •
room temperature, 60V,
200mA, 4 hours
•
4ºC, 125V, 400mA, 1.5
hours
•
Overnight, 30V, 200mA