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Choose the iBlot 3 Western Blot Transfer System when rapid protein transfer, high protein efficiency, reproducibility, and minimal set up and clean up are priorities. Compared to traditional transfer methods, the iBlot 3 system helps deliver efficient and, consistent protein transfer across a broad molecular weight range. This page is for laboratories and core facilities running routine or high-throughput western blot workflows.
Explore the western blot workflow
Efficient, reliable western blot transfer in minutes with the iBlot 3 system
Achieving consistent, high-quality protein transfer is a critical step in western blot workflows. The iBlot Western Blot Transfer System enables rapid, buffer-free protein transfer, enabling fast and reproducible results across experiments.
Designed for performance and convenience, the iBlot 3 system integrates pre-assembled transfer stacks and pre-set programs to streamline setup and minimize variability. This compact, self-contained system supports protein transfer in as few as 3–7 minutes across a broad molecular weight range.
Blot 3 dry transfer is well suited for routine western blot workflows, high-throughput laboratories, and multi-user environments that require consistent, reliable performance.
What is dry western blot transfer?
Dry western blot transfer, also known as dry electrotransfer or dry electroblotting, is a protein transfer method that uses an integrated buffer-containing gel matrix instead of traditional liquid transfer buffer. This compact format enables protein migration under a high electric field without the need for hazardous transfer buffers.
The iBlot 3 Western Blot Transfer System applies this dry transfer technology using pre-assembled transfer stacks that incorporate the membrane and buffer components into a single unit. Pre-set programs control transfer conditions to support consistent protein migration across a broad molecular weight range.
iBlot 3 Western Blot Transfer Device
Gel capacity per run (mini & midi formats) |
Dual transfer stations; 1–4 mini gels or 1–2 midi gels per run |
Transfer time per run |
3–8 min |
Gel size (cm) |
8 x 8 cm, 8 x 13 cm, 13.5 x 10.8 cm |
Gel compatibility |
1.0–1.5 mm thick gels |
Transfer buffer requirements |
Integrated transfer stack (no external liquid buffer required) |
Chemistries |
Tris-glycine; Bis-tris; Tris-acetate; Tricine |
Compatible transfer consumables |
iBlot 3 transfer stacks: low-fluorescence PVDF, PVDF, nitrocellulose (see table below) |
Upgrade program: Are you still using an original iBlot or iBlot 2 device? Learn more about our incentive program to upgrade to an iBlot 3 device.
The iBlot 3 system includes pre-set programs optimized for common protein size ranges. These programs are designed to support efficient electrotransfer without requiring manual adjustment of voltage or current settings.
Use the table below to identify recommended voltage and cooling parameters for common transfer scenarios.
Molecular weight range |
Transfer parameters |
Cooling parameter |
|---|---|---|
Broad range (30–250 kDa) |
25 V, 6 minutes |
Low cooling |
High molecular weight (150–400 kDa) |
30 V, 8 minutes |
No cooling |
Low molecular weight (10–120 kDa) |
25 V, 3 minutes |
Medium cooling |
Transfer stacks for the iBlot 3 system
The iBlot 3 system is compatible with ready-to-use mini and midi transfer stacks available in nitrocellulose (NC), PVDF, and low-fluorescence PVDF formats, supporting a range of chemiluminescent and fluorescent detection workflows.
Low-fluorescence PVDF |
PVDF |
Nitrocellulose |
|---|---|---|
iBlot 3 starter kits
Starter kits are available and include the iBlot 3 Western Blot Transfer Device along with compatible transfer stacks, offering a convenient way to implement dry western blot transfer in your laboratory.
iBlot 3 starter kits |
Western starter kits
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Welcome Pack with iBlot 3 System |
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When do you use iBlot 3 transfer system?
The iBlot 3 Western Blot Transfer System is designed for laboratories that require high transfer efficiency, rapid protein transfer, and reproducible results in a streamlined workflow. By reducing buffer preparation and setup steps, the iBlot 3 system enables protein transfer in minutes—helping teams accelerate western blot workflows.
Dry transfer with the iBlot 3 system is exceptional for:
- Routine western blot experiments requiring fast turnaround
- High-throughput laboratories processing multiple gels per day
- Multi-user environments that benefit from standardized, pre-set methods and two independent transfer stations
- Time-sensitive experiments where rapid transfer is critical
- Laboratories seeking to reduce buffer preparation and hazardous waste cleanup
Because the iBlot 3 system uses pre-optimized programs with customizable settings, it reduces variability and streamlines operation—making it especially well-suited for shared equipment environments and laboratories focused on efficient transfer.
How does iBlot 3 transfer work?
The iBlot 3 Western Blot Transfer System is based on dry western blot transfer concept that integrates the membrane and buffer components into a pre-assembled transfer stack. Each iBlot 3 Transfer Stack consists of a Bottom Stack (copper cathode), a Top Stack (copper anode), and membrane. After protein electrophoresis, the gel is sandwiched between the cathode, the nitrocellulose or PVDF membrane, and the anode, creating a compact environment for controlled protein electrotransfer.
Schematic of dry electroblotting. The schematic illustrates the layered architecture of the iBlot 3 Transfer Stack and the direction of current flow through the gel, membrane, and integrated gel matrices.
Rapid transfer is possible due to the gel matrix design of the iBlot 3 Western Blot Transfer System, which reduces the distance between anode and cathode electrodes and supports higher electric field strength within the compact stack format. Pre-set programs regulate voltage, time, and cooling parameters to maintain controlled electrotransfer conditions.
During transfer:
- The gel matrices at the top and bottom of the stack contain anode and cathode buffers that act as ion reservoirs. This configuration reduces the need for hazardous transfer buffers or soaked filter paper and minimizes handling steps that can lead to inconsistent performance.
- Fresh copper electrodes built into every iBlot transfer stack help improve transfer reproducibility by minimizing variability associated with reused electrodes.
- The shortened distance between the anode and cathode electrodes enables efficient protein migration across a broad molecular weight range while maintaining effective binding to nitrocellulose or PVDF membranes. This design allows the system to increase the transfer speed.
- Integrated cooling enables reliable back-to-back protein transfers by maintaining consistent operating conditions and reducing run-to-run variability.
Transfer stack electrode recycling: The copper electrodes in the iBlot 3 Transfer Stacks can be recycled.
Workflow tip
Western blot of high molecular weight proteins
When routine western blot workflows require rapid turnaround, pair NuPAGE gels with the iBlot 3 Western Blot Transfer System. Dry transfer minimizes setup and buffer handling while delivering high transfer efficiency and reproducible protein transfer in minutes.
Explore NuPAGE Tris-Acetate Gels
iBlot 3 performance
The iBlot 3 system is designed to maximize productivity, helping deliver efficient protein transfer, and provide consistent, reproducible results across experiments.
Maximum productivity
Perform reliable, high-quality protein transfers in as few as 3 minutes. Two independently controlled transfer stations allow you to share the device with your coworker while simultaneously running four mini gels or two midi gels with two different programs. This allows laboratories to complete multiple protein transfers in the time required for a single traditional wet tank transfer, helping increase daily productivity. In addition, you can reduce method setup time by selecting preprogrammed methods from the user-friendly touchscreen interface. Programs can also be customized to fit your needs.
Excellent transfer efficiency
The iBlot 3 Western Blot Transfer System delivers high protein transfer efficiency comparable to or exceeding traditional wet tank and semi-dry transfer methods across a broad range of protein sizes. Unlike traditional wet tank systems that require extended run times and manual buffer preparation, the iBlot 3 system helps deliver rapid, high-efficiency transfer in a compact, self-contained format (Figure 1).
Figure 1. The iBlot 3 system demonstrates better transfer efficiency. (Top) Hsp70: Invitrogen NuPAGE 4–12% Bis-Tris mini gels were loaded with various concentrations of A431 lysate per lane. Immunoprocessing was completed using the Invitrogen Bandmate Western Blot Processor. Thermo Scientific SuperSignal West Dura chemiluminescent substrate was used for detection. Invitrogen iBright FL1500 Imaging System was used for image capture. Local background corrected volume per lysate was plotted for each lane. (Bottom) 4EBP1: Invitrogen Novex 16% Tricine mini gels were loaded with various concentrations of 4EBP1 lysate per lane. Immunoprocessing was completed using the Bandmate western blot processor. Dura chemiluminescent substrate was used for detection. The iBright FL1500 instrument was used for image capture. Local background corrected volume per lysate was plotted for each lane.
Consistent, reproducible results
The pre-assembled, single-use consumables eliminate variability, helping ensure consistent and repeatable results compared with classical methods. Each transfer station delivers comparable protein transfer performance, allowing flexibility to use any station without compromising transfer quality. In addition, the robust design with built-in cooling reduces heat buildup, facilitating run-to-run consistency (Figure 2).
Figure 2. The iBlot 3 device delivers consistent results. A serial dilution of Thermo Scientific HEK293 lysate was loaded onto four Invitrogen Bolt 4–12% Bis Tris plus gels. Invitrogen iBright Prestained Protein Ladder was loaded. Proteins were transferred using the iBlot 3 Western Blot Transfer Device and iBlot 3 Transfer Stacks, Midi, NC. Blots were then blocked with Thermo Scientific Pierce Clear Milk Blocking Buffer and were then probed using primary antibodies raised against EGFR, calreticulin, and p23. Then secondary antibodies AFP-GAR-800, AFP-GAM-488, AF-GAChk-546. Immunoprocessing was completed on the Invitrogen Bandmate Western Blot Processor. Imaging was completed on the Invitrogen iBright FL1500 Imaging System.
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Continue the western blot workflow
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Frequently asked questions
Protein transfer can be completed in as few as 3 minutes for mini gels using optimized pre-set programs. Transfer time depends on gel format and protein molecular weight range.
No. iBlot 3 Transfer Stacks contain integrated gel matrices that function as ion reservoirs, eliminating the need for external liquid transfer buffers or soaked filter paper.
Yes. The iBlot 3 system includes pre-set and customizable programs that support low, medium, and high molecular weight proteins (up to approximately 400 kDa). Optimization may be required depending on protein characteristics and experimental conditions.
Yes. iBlot 3 Transfer Stacks are available with nitrocellulose (0.2 µm) and PVDF (0.2 µm), including low-fluorescence PVDF options for fluorescent western blot applications.
The iBlot 3 system is compatible with commonly used polyacrylamide gel chemistries, including Bis-Tris and Tris-Glycine systems, in mini and midi formats.
Dry electrotransfer uses a pre-assembled stack with an integrated buffer matrix, eliminating external buffer tanks and soaked filter paper. Wet transfer requires liquid buffer and a submerged gel–membrane assembly. Dry transfer typically takes 3–8 minutes, while wet transfer commonly requires 60–120 minutes.
Wet and semi-dry systems use inert electrodes that generate oxygen gas during electrolysis, which can create bubbles between the gel and membrane. Dry electrotransfer uses a copper anode that reduces oxygen generation, helping maintain consistent membrane contact and improving transfer uniformity.
Dry transfer enables faster, more standardized workflows, while wet transfer offers flexibility for protocol customization. Modern dry transfer systems, such as the iBlot 3 system, can deliver high transfer efficiency comparable to or better than traditional wet transfer methods across a broad range of protein sizes, including high molecular weight proteins. The preferred method depends on experimental requirements, workflow priorities, and the level of customization needed.
Western blot educational resources
Access resources to help optimize western blot detection strategies, troubleshoot experiments, and improve data quality.
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