Wet Tank Transfer Systems

Wet transfer is the most common and traditional strategy for western blot transfer due to its efficiency and throughput. We offer three systems for tank transfer that minimize the amount of transfer buffer required.

Transfer membranes  Transfer buffers

Which wet tank transfer system is right for you?

  Invitrogen Mini Blot Module and Mini Gel Tank Invitrogen XCell II Blot Module and SureLock Tank Invitrogen SureLock Tandem Midi Gel Tank with Midi Blot Module
  Mini Blot Module XCell II Blot Module SureLock Tandem Midi Blot Module
Capacity 1 mini gel per blot module; 1-2 blot modules per tank 1 mini gel per blot module; 1-2 blot modules per tank 1 midi gel per blot module, 1-2 blot modules per tank
Transfer buffer requirements 220 mL per blot 800 mL 300 mL per blot
Transfer time 60 min 60-120 min 30 min
Blotting area 9 x 9 cm 9 x 9 cm 9.2 x 14.4 cm
Compatible power supply PowerEase, Owl systems, for other systems use with Novex Power Supply Adapters (Cat. No. ZA10001) PowerEase, Owl systems, for other systems use with Novex Power Supply Adapters (Cat. No. ZA10001) PowerEase or Owl systems
Required equipment Mini Gel Tank XCell SureLock Mini-Cell or XCell II Mini-Cell SureLock Tandem Midi Gel Tank: capacity for up to 2 blot modules

Convenient, reliable western transfer

Two Mini Blot Modules fit into the Mini Gel Tank and are designed to make your western transfers simple and easy to perform.

  • Universal module design—allows modules to fit in either chamber of the tank, simplifying the transfer setup
  • Unique gasket seal—helps prevent buffer leakage so there is no mess during setup of your western transfer
  • 1/2 inch buffer chamber—requires half the typical volume of methanol-based transfer buffers
  • Standard 60 min transfer protocol—accelerates your western workflow so you can get results faster
  • Robust electrodes, sturdy steel plates—for highly efficient and reliable western transfers

Example transfer conditions

Gel type Membrane Voltage Starting current (mA) Ending current (mA) Run time
Bolt Bis-Tris Plus 4-12% (MES) Nitrocellulose 10 160 60 60
PVDF 20 340 130 60
NuPAGE 4-12% Bis-Tris (MES) Nitrocellulose 10 160 60 60
PVDF 20 390 130 60
Novex 4-20% Tris-Glycine (denatured) Nitrocellulose 10 70 50 60
PVDF 20 160 100 60
NuPAGE 3-8% Tris Acetate (denatured) Nitrocellulose 10 150 50 60
PVDF 20 380 130 60
Novex 10-20% Tricine Nitrocellulose 10 70 60 60
PVDF 20 180 150 60

* Current readings represent values when running a single gel, and can vary depending upon the power supply being used.

How-to-videos

How to perform a western wet tank transfer using the Invitrogen Mini Blot Module

Learn how to perform a western blot transfer using the Mini Blot module in a Mini Gel Tank.

How to separate proteins using an Invitrogen precast SDS-PAGE gel

Learn how to separate proteins by using a Invitrogen Bolt Bis-Tris Plus SDS-PAGE precast gel and Mini Gel Tank.

Get started with Mini Bolt Module Welcome Packs

The XCell II Blot Module allows you to easily transfer proteins or nucleic acids from mini-gels to membranes. It fits neatly into the XCell SureLock and XCell II Mini-Cells in place of the gel/buffer core assembly. It requires less than 200 mL of transfer buffer for Western, Southern, and northern transfers. Tough platinized titanium and stainless steel electrodes create a uniform electrical field without clamps or hinged gel holders. Maximum blot size is 9 cm x 9 cm.

Example transfer conditions

Gel type Membrane Transfer buffer Transfer conditions Expected current Run time
Tris-Glycine Nitrocellulose or PVDF Tris-Glycine Transfer Buffer with 20% methanol.

1X Transfer Buffer should be pH 8.3 before addition of SDS or methanol.
25V constant 100 mA 1-2 hr
Tricine Nitrocellulose or PVDF Tris-Glycine Transfer Buffer with 20% methanol.

1X Transfer Buffer should be pH 8.3 before addition of SDS or methanol.
25V constant Start: 100 mA 1-2 hr
Bis-Tris Nitrocellulose or PVDF Bis-Tris transfer buffer with 10% methanol and antioxidant for reduced samples 30 V constant Start: 170 mA End: 100mA 1 hour
Tris-Acetate Nitrocellulose or PVDF Bis-Tris transfer buffer with 10% methanol and antioxidant for reduced samples 30 V constant Start: 200 mA End: 180mA 60
IEF Nitrocellulose or PVDF 0.7% acetic acid, pH 3.0 10 V constant Start: 65-85 mA 60
TBE, TBE-Urea and DAN Retardation Nylon 45 mM Tris, 45 mM boric acid, 1 mM EDTA 30 V constant Start: 360 mA End: 270 mA 1-2 hr

The expected current listed in the table below is for transferring one gel. If you are transferring two gels in the blot module, the expected current will double.

For overnight blotting, transfer at a constant voltage of 10-15 V.

Discover how to use the XCell II Blot Module

Learn how to perform a wet tank transfer using the XCell II Blot Module in the SureLock Tank with this detailed step by step video.

Learn how to run a gel using the XCell SureLock System

Watch this step by step video of how to run a NuPAGE precast get in the SureLock Tank.

Convenient, reliable wet transfer of midi gels

Invitrogen SureLock Tandem Midi Gel Tank with Midi Blot Module

When your western blotting workflow requires higher throughput, Invitrogen midi gels will allow you to load up to 26 samples per gel. You can use the SureLock Tandem Midi Gel Tank to run up to 2 high-performance Invitrogen midi gels and transfer them to membranes in the same tank using the SureLock Tandem Midi Blot Module. Two SureLock Tandem Midi Blot Modules fit into the innovative, dual-purpose SureLock Tandem Midi Gel Tank and are designed for easy, room-temperature gel transfers. Only 300 mL of methanol-containing buffer is required per gel transfer, reducing buffer and hazardous disposal costs.

  • 2-in-1 SureLock Tandem Midi Gel Tank—no need to purchase a separate tank for western transfers
  • Room temperature transfer—save the time, planning, and mess required with other systems to pre-chill buffers, freeze ice packs, or prepare ice baths
  • 300 mL per gel transfer buffer volume—lower transfer buffer volume requirement reduces reagent consumption and hazardous disposal costs
  • Fast and efficient—30-minute transfer without compromising performance
  • Easy to use—convenient blot assembly instructions are printed on the accessory transfer tray
  • Robust electrode plates—for highly efficient and reliable western transfers

 

Get started with a Midi Blot Module Welcome Pack

SureLock Tandem Midi Welcome Pack, Nitrocellulose

SureLock Tandem Midi Welcome Pack

Includes:

  • SureLock Tandem Midi Gel Tank
  • Midi Blot Modules, 2 each
  • SureLock Tandem Midi Pre-cut Membranes and Filters, 0.2 μm, Nitrocellulose
  • SureLock Tandem Transfer Tray
  • Blot Roller
  • Gel knife

Consistent, reliable western blotting results

When used with high-performance Invitrogen precast midi gels, you can depend on the SureLock Tandem Midi Gel Tank and Midi Blot Module to generate consistent, high-throughput western blots. The Invitrogen midi gels offer reproducible high-performance protein separation, resulting in sharp, well-resolved bands in straight lanes, and the SureLock Tandem Midi Blot Module provides dependable uniform transfer of proteins across the gel.

To demonstrate the SureLock Tandem Midi Blot Module’s uniformity of transfer across a membrane, a NuPAGE 4-12% Bis-Tris Midi gel was loaded with recurring dilutions of E. coli lysate ranging from 2-0.25 µg and SeeBlue Plus2 Pre-stained Protein Standard. Electrophoresis was performed and the gel was transferred onto a 0.45 µm PVDF membrane. Results are shown in Figure 1.

A

Data illustrating uniform transfer is achieved using the SureLock Tandem Midi gel Tank

B

Densitometric Signal Linearity versus Protein Load

Figure 1. Uniform transfer using SureLock Tandem Midi Gel Tank. Total protein normalization of transferred proteins shows uniform transfer across the blot. 

A. No-Stain Reagent Labeled PVDF Blot. A NuPAGE 4-12% Bis-Tris Midi gel was loaded with recurring dilutions of E. coli Lysate ranging from 2 to 0.25 µg and SeeBlue Plus2 Pre-stained Protein Standard in lanes 1-2 and 19-20. The gel was electrophoresed using MES running buffer in the SureLock Tandem Midi Gel Tank. Proteins from the gel were transferred onto a 0.45 µm PVDF membranes at 25V constant for 30 minutes using the SureLock Tandem Midi Blot Module. After transfer, the proteins on the membrane were labeled following the No-Stain Protein Labeling Reagent protocol for midi-sized membranes. The image was captured using an iBright Imager with the No-Stain Membrane epi setting (455-485 nm excitation and 565-615 nm emission). As the SeeBlue Plus2 Pre-stained Protein Standard is already labeled, its protein bands are not all susceptible to labeling with No-Stain Reagent, and therefore are not all visible in this image. 

B. Densitometric Signal Linearity versus Protein Load. The densitometric signal intensity was determined for each lane. Technical replicates (n=4) were averaged and plotted to determine the linear regression for the entire concentration range (R2=0.9994). Error bars represent standard deviation. 

Recommended transfer conditions

Recommended transfer conditions for all Invitrogen precast midi gels:
Constant Voltage (V) Time (min)
25 30

Which wet tank transfer system is right for you?

  Invitrogen Mini Blot Module and Mini Gel Tank Invitrogen XCell II Blot Module and SureLock Tank Invitrogen SureLock Tandem Midi Gel Tank with Midi Blot Module
  Mini Blot Module XCell II Blot Module SureLock Tandem Midi Blot Module
Capacity 1 mini gel per blot module; 1-2 blot modules per tank 1 mini gel per blot module; 1-2 blot modules per tank 1 midi gel per blot module, 1-2 blot modules per tank
Transfer buffer requirements 220 mL per blot 800 mL 300 mL per blot
Transfer time 60 min 60-120 min 30 min
Blotting area 9 x 9 cm 9 x 9 cm 9.2 x 14.4 cm
Compatible power supply PowerEase, Owl systems, for other systems use with Novex Power Supply Adapters (Cat. No. ZA10001) PowerEase, Owl systems, for other systems use with Novex Power Supply Adapters (Cat. No. ZA10001) PowerEase or Owl systems
Required equipment Mini Gel Tank XCell SureLock Mini-Cell or XCell II Mini-Cell SureLock Tandem Midi Gel Tank: capacity for up to 2 blot modules

Convenient, reliable western transfer

Two Mini Blot Modules fit into the Mini Gel Tank and are designed to make your western transfers simple and easy to perform.

  • Universal module design—allows modules to fit in either chamber of the tank, simplifying the transfer setup
  • Unique gasket seal—helps prevent buffer leakage so there is no mess during setup of your western transfer
  • 1/2 inch buffer chamber—requires half the typical volume of methanol-based transfer buffers
  • Standard 60 min transfer protocol—accelerates your western workflow so you can get results faster
  • Robust electrodes, sturdy steel plates—for highly efficient and reliable western transfers

Example transfer conditions

Gel type Membrane Voltage Starting current (mA) Ending current (mA) Run time
Bolt Bis-Tris Plus 4-12% (MES) Nitrocellulose 10 160 60 60
PVDF 20 340 130 60
NuPAGE 4-12% Bis-Tris (MES) Nitrocellulose 10 160 60 60
PVDF 20 390 130 60
Novex 4-20% Tris-Glycine (denatured) Nitrocellulose 10 70 50 60
PVDF 20 160 100 60
NuPAGE 3-8% Tris Acetate (denatured) Nitrocellulose 10 150 50 60
PVDF 20 380 130 60
Novex 10-20% Tricine Nitrocellulose 10 70 60 60
PVDF 20 180 150 60

* Current readings represent values when running a single gel, and can vary depending upon the power supply being used.

How-to-videos

How to perform a western wet tank transfer using the Invitrogen Mini Blot Module

Learn how to perform a western blot transfer using the Mini Blot module in a Mini Gel Tank.

How to separate proteins using an Invitrogen precast SDS-PAGE gel

Learn how to separate proteins by using a Invitrogen Bolt Bis-Tris Plus SDS-PAGE precast gel and Mini Gel Tank.

Get started with Mini Bolt Module Welcome Packs

The XCell II Blot Module allows you to easily transfer proteins or nucleic acids from mini-gels to membranes. It fits neatly into the XCell SureLock and XCell II Mini-Cells in place of the gel/buffer core assembly. It requires less than 200 mL of transfer buffer for Western, Southern, and northern transfers. Tough platinized titanium and stainless steel electrodes create a uniform electrical field without clamps or hinged gel holders. Maximum blot size is 9 cm x 9 cm.

Example transfer conditions

Gel type Membrane Transfer buffer Transfer conditions Expected current Run time
Tris-Glycine Nitrocellulose or PVDF Tris-Glycine Transfer Buffer with 20% methanol.

1X Transfer Buffer should be pH 8.3 before addition of SDS or methanol.
25V constant 100 mA 1-2 hr
Tricine Nitrocellulose or PVDF Tris-Glycine Transfer Buffer with 20% methanol.

1X Transfer Buffer should be pH 8.3 before addition of SDS or methanol.
25V constant Start: 100 mA 1-2 hr
Bis-Tris Nitrocellulose or PVDF Bis-Tris transfer buffer with 10% methanol and antioxidant for reduced samples 30 V constant Start: 170 mA End: 100mA 1 hour
Tris-Acetate Nitrocellulose or PVDF Bis-Tris transfer buffer with 10% methanol and antioxidant for reduced samples 30 V constant Start: 200 mA End: 180mA 60
IEF Nitrocellulose or PVDF 0.7% acetic acid, pH 3.0 10 V constant Start: 65-85 mA 60
TBE, TBE-Urea and DAN Retardation Nylon 45 mM Tris, 45 mM boric acid, 1 mM EDTA 30 V constant Start: 360 mA End: 270 mA 1-2 hr

The expected current listed in the table below is for transferring one gel. If you are transferring two gels in the blot module, the expected current will double.

For overnight blotting, transfer at a constant voltage of 10-15 V.

Discover how to use the XCell II Blot Module

Learn how to perform a wet tank transfer using the XCell II Blot Module in the SureLock Tank with this detailed step by step video.

Learn how to run a gel using the XCell SureLock System

Watch this step by step video of how to run a NuPAGE precast get in the SureLock Tank.

Convenient, reliable wet transfer of midi gels

Invitrogen SureLock Tandem Midi Gel Tank with Midi Blot Module

When your western blotting workflow requires higher throughput, Invitrogen midi gels will allow you to load up to 26 samples per gel. You can use the SureLock Tandem Midi Gel Tank to run up to 2 high-performance Invitrogen midi gels and transfer them to membranes in the same tank using the SureLock Tandem Midi Blot Module. Two SureLock Tandem Midi Blot Modules fit into the innovative, dual-purpose SureLock Tandem Midi Gel Tank and are designed for easy, room-temperature gel transfers. Only 300 mL of methanol-containing buffer is required per gel transfer, reducing buffer and hazardous disposal costs.

  • 2-in-1 SureLock Tandem Midi Gel Tank—no need to purchase a separate tank for western transfers
  • Room temperature transfer—save the time, planning, and mess required with other systems to pre-chill buffers, freeze ice packs, or prepare ice baths
  • 300 mL per gel transfer buffer volume—lower transfer buffer volume requirement reduces reagent consumption and hazardous disposal costs
  • Fast and efficient—30-minute transfer without compromising performance
  • Easy to use—convenient blot assembly instructions are printed on the accessory transfer tray
  • Robust electrode plates—for highly efficient and reliable western transfers

 

Get started with a Midi Blot Module Welcome Pack

SureLock Tandem Midi Welcome Pack, Nitrocellulose

SureLock Tandem Midi Welcome Pack

Includes:

  • SureLock Tandem Midi Gel Tank
  • Midi Blot Modules, 2 each
  • SureLock Tandem Midi Pre-cut Membranes and Filters, 0.2 μm, Nitrocellulose
  • SureLock Tandem Transfer Tray
  • Blot Roller
  • Gel knife

Consistent, reliable western blotting results

When used with high-performance Invitrogen precast midi gels, you can depend on the SureLock Tandem Midi Gel Tank and Midi Blot Module to generate consistent, high-throughput western blots. The Invitrogen midi gels offer reproducible high-performance protein separation, resulting in sharp, well-resolved bands in straight lanes, and the SureLock Tandem Midi Blot Module provides dependable uniform transfer of proteins across the gel.

To demonstrate the SureLock Tandem Midi Blot Module’s uniformity of transfer across a membrane, a NuPAGE 4-12% Bis-Tris Midi gel was loaded with recurring dilutions of E. coli lysate ranging from 2-0.25 µg and SeeBlue Plus2 Pre-stained Protein Standard. Electrophoresis was performed and the gel was transferred onto a 0.45 µm PVDF membrane. Results are shown in Figure 1.

A

Data illustrating uniform transfer is achieved using the SureLock Tandem Midi gel Tank

B

Densitometric Signal Linearity versus Protein Load

Figure 1. Uniform transfer using SureLock Tandem Midi Gel Tank. Total protein normalization of transferred proteins shows uniform transfer across the blot. 

A. No-Stain Reagent Labeled PVDF Blot. A NuPAGE 4-12% Bis-Tris Midi gel was loaded with recurring dilutions of E. coli Lysate ranging from 2 to 0.25 µg and SeeBlue Plus2 Pre-stained Protein Standard in lanes 1-2 and 19-20. The gel was electrophoresed using MES running buffer in the SureLock Tandem Midi Gel Tank. Proteins from the gel were transferred onto a 0.45 µm PVDF membranes at 25V constant for 30 minutes using the SureLock Tandem Midi Blot Module. After transfer, the proteins on the membrane were labeled following the No-Stain Protein Labeling Reagent protocol for midi-sized membranes. The image was captured using an iBright Imager with the No-Stain Membrane epi setting (455-485 nm excitation and 565-615 nm emission). As the SeeBlue Plus2 Pre-stained Protein Standard is already labeled, its protein bands are not all susceptible to labeling with No-Stain Reagent, and therefore are not all visible in this image. 

B. Densitometric Signal Linearity versus Protein Load. The densitometric signal intensity was determined for each lane. Technical replicates (n=4) were averaged and plotted to determine the linear regression for the entire concentration range (R2=0.9994). Error bars represent standard deviation. 

Recommended transfer conditions

Recommended transfer conditions for all Invitrogen precast midi gels:
Constant Voltage (V) Time (min)
25 30

Resources