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Transfer Buffers

AquaBlot™ 10×High Efficiency Transfer Buffer is a 10 x transfer buffer with higher transfer efficiency than the conventional Towbin (tris-glycine) buffers. Use it after 10-fold dilution with deionized water. Addition of methanol is not required during preparation.

* Methanol is necessary for hydrophilization of the PVDF membrane.

Features

  • Higher transfer efficiency than conventional products
  • Simply change the transfer buffer
  • No methanol needed

Comparison with conventional products

1. Constant voltage condition

Method

After 60 minutes of (semi-dry) transfer at 25 V constant voltage, an appropriate exposure time for comparison of luminescence intensity was selected, and the luminescence intensity value of each protein was read and expressed as a relative value. After 60 minutes of (semi-dry) transfer at 25 V constant voltage, an appropriate exposure time for comparison of luminescence intensity was selected, and the luminescence intensity value of each protein was read and expressed as a relative value.
After the signals from low molecular weight proteins were read, the PVDF membrane was excised to measure the luminescence intensity of high molecular weight proteins, which require prolonged exposure, separately.

Results

Under constant voltage conditions, AquaBlot™ 10 x High Efficiency Transfer Buffer exhibited higher transfer efficiency than Towbin and Towbin (SDS) Buffers, in both the high and low molecular weight regions.

Conditions

    • AquaBlot™
      AquaBlot™ 10 x High Efficiency Transfer Buffer (10-fold dilution)
    • Towbin buffer
      25 mmol/L Tris, 192 mmol/L Glycine with 10 v/v% Methanol
    • Towbin buffer(SDS)
      25 mmol/lL Tris, 192 mmol/L Glycine, 0.05 w/v% SDS with 10 v/v% Methanol
    • Gel
      SuperSep™Ace, 10-20%, 17 well (Product Number: 198-15041)
    • Membrane
      ClearTrans® SP PVDF Membrane, Hydrophobic, 0.2μm (Product Number: 033-22453)
    • Chemiluminescence reagent
      ImmunoStar® Zeta (Product Number: 295-72404)
2. Constant current condition

Method

After 60 minutes of (semi-dry) transfer at a constant current of 1.2 mA/cm2, an appropriate exposure time for comparison of luminescence intensity was selected, and the luminescence intensity value of each protein was read and expressed as a relative value. After the signals from low molecular weight proteins were read, the PVDF membrane was excised to measure the luminescence intensity of high molecular weight proteins, which require prolonged exposure, separately.

Results

Under constant current conditions, AquaBlot™ 10 x High Efficiency Transfer Buffer exhibited higher transfer efficiency than Towbin and Towbin (SDS) Buffers, in both high and low molecular weight regions.

Conditions

    • AquaBlot™
      AquaBlot™ 10 x High Efficiency Transfer Buffer (10-fold dilution)
    • Towbin buffer
      25 mmol/L Tris, 192 mmol/L Glycine with 10 v/v% Methanol
    • Towbin buffer(SDS)
      25 mmol/lL Tris, 192 mmol/L Glycine, 0.05 w/v% SDS with 10 v/v% Methanol
    • Gel
      SuperSep™ Ace 10–20%, 17-well (Product Number: 198-15041)
    • Membrane
      ClearTrans® SP PVDF Membrane, Hydrophobic, 0.2μm (Product Number: 033-22453)
    • Chemiluminescence reagent
      ImmunoStar® Zeta (Product Number: 295-72404)

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