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Choose wet transfer systems based on throughput, gel compatibility, and transfer requirements. Compare wet tank blot modules and configurations to support flexible transfer conditions, high-capacity sample processing, and reliable protein transfer across applications. This page is for laboratories prioritizing throughput, flexibility, and long-term operating value.
Explore the western blot workflow
Reliable, flexible wet transfer for western blot workflows
Wet transfer is a widely used, traditional method for protein transfer in western blot workflows, offering reliable performance and the flexibility to tailor transfer conditions to specific experimental needs. As a trusted, widely used approach, it remains a standard choice for laboratories working with a wide range of proteins and established protocols.
Invitrogen wet transfer systems support high-capacity processing (up to 52 samples simultaneously), enabling multiple gels to be transferred in parallel while maintaining consistent results across experiments. Designed for compatibility across a wide range of gel chemistries and membrane types, these systems support adaptable workflows and reproducible protein transfer.
Their flexibility allows researchers to tailor transfer conditions based on protein size, gel chemistry, and experimental requirements, making wet transfer a widely used approach for established western blot workflows.
What is western blot wet transfer?
Wet transfer (also known as wet tank transfer) is a traditional electroblotting method used to transfer proteins from an SDS-PAGE gel to a membrane (PVDF or nitrocellulose) using an electric field in a buffer-filled tank. This method typically uses larger buffer volumes and greater electrode spacing compared to semi-dry or dry transfer approaches, enabling controlled protein migration and flexibility in adjusting transfer conditions such as buffer composition, voltage, and transfer time.
Invitrogen wet transfer systems include the Mini Blot Module, XCell II Blot Module, and SureLock™ Tandem Midi Blot Module. These systems incorporate reduced buffer chamber designs to help minimize methanol use and associated waste while maintaining consistent transfer performance. Flexible buffer composition and consumable configurations allow researchers to optimize transfer conditions while managing reagent use and cost.
Wet transfer is widely used for established western blot workflows, particularly in laboratories processing multiple gels due to their robust performance, adaptability, and consistent results.
Which wet transfer system is right for you?
Invitrogen wet transfer systems support flexible western blot workflows across mini and midi gel formats.
Use the table below to compare wet transfer system capabilities.
|
 Invitrogen Mini Blot Module |
 Invitrogen XCell II Blot Module |
|
|---|---|---|---|
Supported gel format |
Mini gels |
Mini gels |
Midi gels |
Gel capacity per run (mini & midi formats) |
1 mini gel per blot module (1–2 blot modules per tank) |
1–2 mini gel per blot module (1 blot module per tank) |
1 midi gel per blot module (1–2 blot modules per tank) |
Transfer time per run |
60 min |
60–120 min |
30 min |
Blotting area |
9 x 9 cm |
9 x 9 cm |
9.2 x 14.4 cm |
Buffer volume |
220 mL per blot module |
200 mL per blot module |
300 mL per blot module |
Compatible tank |
Need a complete module and tank set? Mini Gel Tank and Blot Module Set, XCell SureLock Mini-Cell and XCell II Blot Module , or see welcome packs below.
Mini blot module
Use the Invitrogen Mini Blot Module when your workflow uses the Mini Gel Tank for a simplified setup, reduced buffer consumption, and flexibility for routine workflows.
Key advantages:
- Universal module design—fits in either chamber of the tank, simplifying the transfer setup
- Secure gasket seal—helps prevent buffer leakage during western transfer setup
- Less than 2-inch buffer chamber—reduced volume of methanol-based transfer buffers
- Standard 60 min protocol—accelerates your western workflow, so you can get results fast
- Robust electrodes and steel plates—helps deliver efficient and reliable western transfers
How it fits into your workflow
The mini blot module is designed for routine mini-gel transfers. After electrophoresis, the gel and membrane are assembled into the transfer cassette and inserted into the Mini Gel Tank. Two modules can operate simultaneously within the same tank, supporting efficient processing of multiple samples with a standard 60-minute protocol.
XCell II blot module
Choose the Invitrogen XCell II Blot Module when your workflow uses the XCell SureLock system, enabling easy workflow continuity without additional equipment or buffer-intensive setup.
Key advantages:
- Integrated system compatibility—fits directly into compatible XCell systems
- 200 mL transfer buffer requirement—reduce buffer waste for western, southern, and northern transfers
- Uniform electrical field with platinized titanium and stainless-steel electrodes—no clamps or hinged gel holders required
How it Integrates with XCell systems
The XCell II blot module replaces the gel/buffer core assembly in the XCell SureLock system. Following electrophoresis, the gel is transferred directly within the same platform using 200 mL of transfer buffer, enabling easy workflow continuity without additional tank systems.
How do I use the XCell II Blot Module in an XCell SureLock system?
Efficient protein transfer is a critical step in western blot workflows. In this video, you’ll learn how to transfer proteins from polyacrylamide mini gels to nitrocellulose or PVDF membranes using the XCell II Blot Module in a SureLock tank.
You’ll learn how to:
- Prepare the required materials to run a wet transfer
- Assemble the gel-membrane transfer sandwich
- Install the XCell II Blot Module into the XCell SureLock system
- Add the appropriate buffers and run a wet protein transfer
- Disassemble the blot module and handle the membrane for downstream western blot detection
SureLock tandem midi blot module
Select the Invitrogen SureLock Tandem Midi Blot Module when your workflow uses the SureLock Tandem Midi Gel Tank and requires higher throughput or midi-gel capacity.
Key advantages:
- Room temperature transfer—no pre-chilling or ice baths, increasing efficiency and reducing mess
- Reduced buffer volume (~300 mL per gel)—lowers reagent consumption and disposal costs
- Fast and efficient—30-minute transfer with consistent performance
- Dual midi-gel capacity—supports higher-throughput workflows
- Printed assembly instructions on tray—easy-to-use and convenient
- Robust electrode plates—for highly efficient and reliable western transfers
How it supports higher throughput
The SureLock Tandem midi system allows you to run and transfer up to two midi gels within the same dual-purpose tank—allowing up to 26 samples per gel. After electrophoresis, the blot module is inserted into the tank for a fast 30-minute, room-temperature transfer, removing the need for pre-chilling buffers or separate equipment.
Demonstrated uniform transfer performance
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 offers 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 Plus 2 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.
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 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 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). (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.
Wet transfer welcome packs
Whether you're implementing wet transfer for the first time or expanding throughput, wet transfer welcome packs include essential components needed to get started quickly. Choose between mini and midi configurations based on gel capacity requirements and select nitrocellulose or PVDF membrane options to match your downstream detection workflow.
Each pack includes the required tank and blot modules, transfer buffer, and membrane/filter components to streamline setup and reduce preparation time.
 |
 |
 |
|
|---|---|---|---|
| Bolt Western Pack A (Nitrocellulose) | Bolt Western Pack B (PVDF) | ||
Each pack contains:
|
Each pack contains:
|
Contains:
|
Contains:
|
| Required tank: Mini Gel Tank | |||
Recommended transfer conditions
Recommended transfer conditions vary depending on protein size, gel chemistry, and experimental goals. The conditions in the table below serve as starting points and may require optimization to achieve optimal transfer efficiency for your experimental needs.
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.
Gel type |
Membrane |
Transfer buffer |
Transfer conditions |
Expected current |
Run time |
|---|---|---|---|---|---|
Tris-Glycine |
Nitrocellulose or PVDF |
Tris-Glycine Transfer Buffer with 20% methanol. |
25 V constant |
100 mA |
1–2 hr |
Tricine |
Nitrocellulose or PVDF |
Tris-Glycine Transfer Buffer with 20% methanol. |
25 V 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: 100 mA |
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: 180 mA |
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.
Recommended transfer conditions for Invitrogen precast midi gels:
Constant voltage (V) |
Time (min) |
25 |
30 |
Ordering information
Continue the western blot workflow
Explore additional solutions that support each step of the western blot workflow from protein transfer to data analysis:
Frequently asked questions
Wet transfer uses an electric field to move negatively charged proteins from a polyacrylamide gel onto a membrane submerged in transfer buffer. The gel–membrane sandwich is placed into a cassette, submerged in buffer within the transfer tank, and exposed to a controlled electrical current. Proteins migrate out of the gel and bind to the membrane for downstream detection.
Transfer time typically ranges from 30 minutes to 2 hours, depending on protein size, gel percentage, and voltage conditions. Additionally, overnight transfers are possible with a modified reduced voltage protocol.
Wet tank transfer systems are compatible with PVDF and nitrocellulose membranes.
Yes. Wet tank transfer systems allow simultaneous transfer of multiple gels depending on blot module capacity.
Wet transfer is preferred when flexibility and control over transfer conditions are required, allowing manual adjustment of buffer composition, voltage, and transfer time. Semi-dry transfer offers a balance of speed and flexibility with reduced buffer usage. Dry transfer systems help provide the fastest set-up and transfer with minimal buffer handling and pre-set or customizable programs, supporting consistent and streamlined workflows. The best method ultimately depends on experimental needs, workflow priorities, and the level of customization required.
Western blot educational resources
Access resources to help optimize western blot detection strategies, troubleshoot experiments, and improve data quality.
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