Bio-Rad Trans-Blot® Cell - Manual (Page 11)

2.3 Acidic Transfers
If transferring under acidic conditions, switch the gel and membrane in the set up instruc-
tions. This will place the membrane on the cathode side of the gel. Under acidic conditions,
proteins will transfer in the opposite direction going toward the negative cathode. Do not
reverse the electrodes themselves. This will cause damage to the instrument.
Section 3
Transfer Conditions
3.1 General Guidelines to Transfer Buffers
and Running Conditions
Tables 3.1 to 3.4 provide guidelines for power conditions using different buffers. Power
conditions are provided for various run times. Where multiple conditions are displayed, the
higher the voltage, the less time required for the run. Note all “requires cooling” warnings.
Table 3.1. SDS-PAGE Gels
Standard Field Standard Field High Intensity Field
Overnight 8 cm electrode distance 4 cm electrode distance
❄
C C C CO O O OO O O OLLLLIIIIN N N NG G G G R R R REE E EQ Q Q QU U U UIIIIR R R REE E ED D D D
Buffer A or B Buffer C Buffer A or B Buffer C Buffer A or B Buffer C
Trans-Blot cell with 30 V, 0.1 A 10 V, 0.1 A 60 V, 0.21 A 20 V, 0.25 A 100–200 V 30–60 V
wire electrodes 0.3–0.8 A 0.4–1.0 A
Overnight 5 hours approximately 30 minutes–4 hours ❄
Buffer A or B Buffer C Buffer A or B Buffer C Buffer A or B Buffer C
Trans-Blot cell with 10 V 5 V 100–150 V 30–50 V 50–100 V 20–40 V
plate electrodes 0.1 A 0.1 A 1–1.5 A 1–1.6 A 0.7–1.6 A 0.7–1.6 A
Overnight 1–5 hours ❄ 30 minutes–1 hour ❄
Buffer A: 25 mM Tris, pH 8.3, 192 mM glycine, with or without 20% MEOH and .025%–0.1% SDS.
B: 48 mM Tris, pH 9.2, 39 mM glycine, with or without 20% MEOH and .025%–0.1% SDS.
C: 10 mM NaHCO
3
, 3 mM NaCO
3
, pH 9.9, with or without 20% MEOH and .025%–0.1% SDS.
8