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3 acidic transfers – Bio-Rad Trans-Blot® Cell User Manual

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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.

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