C.B.S. Scientific TTGE-2401 - Manual (Page 9)

www.cbsscientific.com 9
GEL BLOTTING
Follow general guidelines as indicated for respective blotting units. As a general recommendation, equilibrate
gels (after running) with the diluted transfer buffer for 5 to 10 minutes prior transfer. Clear PAGE™ Transfer
Buffer 10x concentrate contains 0.25 M Tris (base), 1.92M Glycine, 1% SDS.
For C.B.S. Scientific’s EBU-204, EBX-700, DCX-700, EBU-4000 and EBU-6000 Blotter Buffers:
Component EBX-700
EBU-204
Tank Blotters
DCX-700
Blotter
EBU-4000 and EBU-6000
Semi-Dry Blotters
ClearPAGE™ Transfer Buffer
10X or 20X dilution
(cat. FB82500)
50ml
(1:20 dilution)
100ml
(1:10 dilution)
10ml
(1:10 dilution)
Methanol 200ml 200ml 20ml (for PVDF membrane:
reduce 50%)
Ultrapure water 770ml 720ml 72 ml (PVDF: add water for less
MeOH)
Typical Blotting
conditions
EBX-700, EBU-204
Tank Blotters
DCX-700 or
Quadra
EBU-4000 & EBU-6000
Semi-Dry Blotting
Power Supply Setting 50-60V constant 200V constant 25 Volts
Blot time 2-4 hours with stirring. Ex-
change cooling blocks after
1.5 hours
1.5 - 2.0 hours with
stirring, cooling
blocks
30-60 minutes
Expected current 250-300mA 180mA / 1 gel
220mA / 2 gels
440mA/ 4 gels
Initial 200-300mA / Final 60-
100mA
Refer to figure below to assemble blotting stack. With cassette wide open assemble components on black
side in the following order: buffer saturated sponge pad, gel equilabrated in transfer buffer, buffer saturated
transfer membrane, then buffer saturated blotting paper. Smooth with gloved finger or roll with glass rod to
be sure no bubbles exist between the gel and the transfer membrane.
red side
high molecular weight
bands at this end
blotting paper
gel
foam pad
black side
membrane
3