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Explore iBind Western Blot System capabilities for passive-flow antibody incubation and washing to support simplified membrane processing, minimal setup, and operation without electricity, vacuum pumps, or controllers.
Explore the western blot workflow
iBind western blot system for hands-free antibody incubation and washing
Western blot processing includes membrane blocking, antibody incubation, and washing steps prior to detection.
The iBind western blot system simplifies these steps by enabling hands-free membrane processing without pumps, power, or programming. Using sequential lateral flow technology, reagents are added across the membrane in a controlled sequence, providing results in about 2 hours.
Designed for ease of use and minimal setup, the iBind system supports consistent, reproducible western blot processing in a compact, self-contained format.
What is the iBind western blot system?
The iBind western blot system is a hands-free western blot system that performs membrane blocking, antibody incubation, and washing using sequential lateral flow technology. Unlike programmable systems, iBind systems use passive flow to move reagents across the membrane without the need for power, pumps, or external equipment.
The iBind systems control reagent flow supporting consistent, reproducible membrane processing. In addition, iBind PLUS and iBind Flex PLUS solution kits are formulated to support efficient antibody-target interactions and reliable signal detection.
iBind western blot systems
The iBind western blot systems consist of the iBind Western Device, iBind sequential lateral flow cards, and iBind solution kits for chemiluminescent or fluorescent detection. Two throughput options are available:
- iBind western blot system—processes one mini blot
- iBind Flex western blot system—processes two mini blots, one midi blot or six vertically cut strips
The iBind system is well suited for:
- Simple, hands-free western blot processing
- Workflows prioritizing minimal setup and ease of use
- Consistent membrane processing with reduced user variability
- Efficient workflows that reduce hands-on time and antibody consumption
Tip: Choose iBind for hands-free membrane processing with minimal setup.
Which iBind western blot system is right for you?
Two iBind western systems are available: the original iBind Western Blot Device, and the iBind Flex Western Blot Device. Setup requires only approximately 15 minutes, with no additional hands-on steps until the blot has been processed and is ready for the detection step.
Use this table to compare iBind system formats and processing capabilities.
Features |
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Mini blot (single) |
Yes |
Yes |
Mini blot (dual) |
No |
Yes |
Midi blot |
No |
Yes |
Vertically cut strips |
No |
Yes |
Format flexibility |
Single mini blot workflows |
Multiple formats and sample types |
For workflows requiring programmable control or higher throughput, consider the Bandmate western blot processor.
iBind starter kits
Our starter kits include everything you need to start processing your western blots using the iBind western systems.
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Use the new iBind Starter Kit and iBind Flex Starter Kit with iBind Plus.
iBind and iBind Flex solution kits
iBind solution kits are available for either chemiluminescent or fluorescent detection.
For chemiluminescent, chromogenic, colorimetric detection
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For fluorescent detection, including infrared detection and dual wavelength semi-qualitative analysis |
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10 mini blot reactions |
10 midi or 20 mini blot reactions |
10 mini blot reactions |
10 midi or 20 mini blot reactions |
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Includes:
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Includes:
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iBind and iBind Flex cards
The iBind western systems utilize specialized iBind Cards for each run and consist of a flow region and a stack. The card is a matrix optimized for homogenous flow and direct contact of reagents across the western blot membrane. iBind Cards are available in two sizes for the iBind system or iBind Flex system.
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iBind Flex well formats
The iBind Flex system comes with interchangeable wells, which allows you to run multiple membrane formats and even run different primary and secondary antibody conditions in the same device at the same time.
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iBind workflow advantages
- Hands-free membrane processing—load reagents and membrane at the start; processing proceeds without further input
- Consistent, reproducible results—controlled reagent flow helps reduce blot-to-blot variability
- Easy setup—no programming, pumps, or external equipment required
- Faster blot processing—reduced processing from overnight or several hours to approximately 2.5 hours
- Efficient antibody usage—low reagent volumes help reduce primary antibody consumption by 80% (2 mL vs. 10 mL)
- Compact and flexible design—compatible with PVDF, nitrocellulose, and multiple detection methods
- No electricity required—self-contained operation without external power
The iBind western system uses a self-contained format that simplifies membrane processing while maintaining consistent incubation and washing conditions.
How does iBind system western blot processing work?
The iBind western system performs membrane blocking, antibody incubation, and washing using sequential lateral flow technology to help deliver hands-free western blot processing.
Simplified and efficient workflow
iBind western blot system’s self-contained design reduces setup complexity and supports consistent processing across different users and experiments. Its hands-free membrane processing allows all solutions to be prepared and loaded in the device at the start of the workflow, with no additional user intervention required during incubation and washing steps. All subsequent steps proceed automatically and uninterrupted by sequential lateral flow technology (SLF), e.g., simple capillary action—no electricity, pumps, or external instrumentation. The iBind western systems significantly reduce the time and effort involved compared to traditional western blotting processes (Figure 1).
Figure 1. Time savings of automated western blotting processing with the iBind western blot system.
At-a-glance workflow:
Hands-free automated membrane processing in 3 steps:
- Load reagents and membrane
- Start the run (no programming required)
- Retrieve membrane ready for detection
How sequential lateral flow works
The iBind western blot system achieves automated blot processing without the use of a complex, high-maintenance fluidics system. Instead, it relies on sequential lateral flow (SLF). Similar to rapid diagnostic tests, SLF is based on the ability of a liquid to move through paper or a specialized matrix via capillary action.
- Reagents are added to designated wells at the start of the experiment
- Solutions flow across the membrane in a controlled sequence
- Each step (blocking → antibody incubation → washing) occurs automatically
- Flow is driven without pumps, tubing, or external power
This design enables consistent reagent delivery and reduces the need for manual timing or handling during processing (Figure 2).
Figure 2. Illustration of how sequential lateral flow technology enables hands-free western blot membrane processing with the iBind western blot system.
How does sequential lateral flow technology work?
Sequential lateral flow (SLF) technology enables hands-free western blot membrane processing by moving reagents across the membrane in a controlled sequence without the need for pumps, tubing, or external power. In this video, you’ll learn how the iBind Western Device automates membrane processing and supports consistent, reproducible results with minimal setup.
You’ll learn how to:
- Understand how SLF moves reagents across the membrane using capillary action
- See how blocking, antibody incubation, and washing steps occur in sequence
- Visualize how consistent reagent delivery supports reliable western blot results
Reagents are prepared using standard western blot protocols and loaded into the device in a defined sequence.
- Prepare and wet the iBind or iBind Flex card before placing the membrane into the system
- Ensure proper membrane placement and contact with the processing matrix
- Follow the recommended reagent loading order
- Avoid disturbing the system during operation
Once assembled, processing proceeds automatically without further user input, enabling a simple, load-and-go workflow.
Tips for optimal results:
- Use appropriate antibody concentrations
- Inspect cards for damage or excessive wrinkling before use
- Ensure firm membrane contact with the processing matrix
- Avoid disturbing the system during operation
iBind performance
The iBind is designed to streamline membrane processing while supporting consistent, reproducible results across experiments.
Sequential lateral flow (SLF) helps deliver antibody and wash solutions across the membrane in a controlled and uniform manner. This enables efficient target binding and rapid removal of unbound antibody, reducing reagent volumes and supporting higher secondary antibody concentrations without increasing background. SLF supports consistent incubation and washing conditions, reducing variability between experiments.
The iBind western blot system uses low reagent volumes, only requiring 2 mL of antibody solution per mini blot compared to larger volumes used in manual workflows. This can significantly reduce overall antibody consumption (Figure 3–5).
Primary antibodies can represent a substantial portion of western blot experiment costs. By reducing required volumes, the iBind system helps support more efficient reagent use without increasing workflow complexity.
Figure 3. Use 5X less primary antibody per experiment with the iBind PLUS Solution Kit. A dilution series (2-Fold) of cell lysate (5 µg lysate start) was separated and then transferred to nitrocellulose membranes using the iBlot 3 Transfer Device. Membranes were then incubated with primary and secondary antibodies for the detection of 3 different protein targets (4EBP1, Caspase 7, and AKT) using either the iBind western blot system with iBind PLUS Solution Kit or standard western processing of manual blocking, washing and antibody incubation. The same primary antibody concentration of 1:1000 was used for all western blots, while a 10X higher concentration of secondary antibody was used for iBind blot processing (1:10,000) vs. manual western blot processing (1:100,000).
In addition to reduced antibody volume, the antibody diluent in the iBind PLUS and iBind Flex PLUS solution kits (for chemiluminescent detection) is formulated to support efficient antibody-target interactions. When used with standard antibody dilutions, these kits can enable increased signal detection compared to standard overnight western blot workflows (Figure 4). As a result, primary antibody concentration can often be further reduced to generate similar detection sensitivity as manual methods while maintaining reliable detection across many protein targets (Figure 5).
Figure 4. iBind western blot systems with iBind PLUS solution kits demonstrate stronger signal detection to manual western blot processing under equivalent exposure times. For each target, a dilution series (2-Fold) of cell lysate (5 µg lysate start) was separated and then transferred to nitrocellulose membranes using the iBlot 3 Transfer Device. Membranes were then incubated with primary and secondary antibodies using either iBind western blot systems and iBind PLUS Solution Kit (left) or by manual processing (right) for the detection of 10 different protein targets. Blots processed with the iBind western systems and iBind PLUS Solution Kit used 2 mL of diluted primary antibody (1:1,000) and 2 mL of diluted secondary-HRP antibody (1:10,000). Manually processed blots were immunoprobed in 10 mL of primary antibody diluted 1:1000 in blocker for 16 hours with rocking, washed with TBST (3 x 10 mL for 5 minutes each), incubated in 10 mL of 1:100,000 diluted HRP-labeled secondary antibody for 1 hour, followed by final washing in TBST (5x10 mL for 5 minutes each).
Figure 5. Primary antibody concentration can be significantly reduced without compromising performance. A431 lysate starting at 10 µg was two-fold serial diluted, separated on Novex 4–20% Tris-Glycine gels, and transferred on NC membrane using the iBlot 3 Western Blot Transfer Device. Membranes were then processed with primary antibodies for 3 targets (AKT, Ezrin, and MEK1) using AKT Pan Monoclonal Antibody, Ezrin Monoclonal Antibody, or MEK1 Monoclonal Antibody, respectively, at varying concentrations as indicated using the iBind western blot systems with iBind PLUS Solution Kit, followed by incubation with secondary-HRP antibody diluted at 1:10,000 as recommended for the iBind protocol. Manual western blotting was processed using Pierce Clear Milk Blocking Buffer for 1 hour, followed by the same primary antibodies as used in iBind western system for each respective target at a 1:1,000 in Pierce Clear Milk Blocking Buffer and incubated overnight at 4°C. Secondary-HRP antibody was diluted 1:100,000 for each blot. All blots were incubated with SuperSignal West Dura substrate and imaged under optimal exposure times for each blot using the iBright FL1500 Imaging System.
When combining the iBind Flex Device along with chemiluminescent detection kit (iBind Flex PLUS solution kit—Figures 3–5) or fluorescent detection kit (iBind Flex FD solution kit), multiplex western blot workflows can achieve consistent detection with sensitivity comparable to manual processing (Figure 6).
Figure 6. Similar or greater detection sensitivity in multiplex experiments. NuPAGE Bis-Tris 4–12% precast gels were loaded with dilution series (2-Fold) of cell lysate and then separated and transferred to nitrocellulose membranes using the iBlot 3 Transfer Device. Membranes were then incubated with primary and secondary antibodies using either iBind Flex Western Blot System and iBind Flex Fluorescent Detection (FD) Solution Kit (top) or by manual processing (bottom) for the multiplex detection of A) PDI and Cyclophilin B (left) or B) Hsp90 and p23 (right). Blots processed with the iBind Flex Western Device and iBind Flex Fluorescent Detection (FD) Solution Kit used 2 mL of diluted primary antibody (1:500) and 2 mL of diluted fluorescently conjugated secondary antibody (1:1,000). Manually processed blots were immunoprobed with diluted primary antibodies (1:1,000 in 10 mL for each respective target), followed by Alex Flour Plus-conjugated secondary antibodies (1:5,000 in 10 mL) corresponding to each primary antibody. Each set of blots were exposed and imaged together using the iBright FL1500 Imaging System.
Increase productivity with iBind Flex Device as part of the iWestern 4-hour workflow
The iBind Flex Western Device supports flexible membrane formats and enables larger or parallel experiments within a single workflow. Interchangeable well inserts allow processing of multiple membrane formats or different antibody conditions simultaneously, supporting up to two mini blots, one midi blot, or vertically cut membrane strips in a single run.
When integrated into the broader iWestern 4-hour workflow, iBind Flex enables streamlined, end-to-end western blot processing—from electrophoresis and transfer through detection and imaging—helping researchers complete single to multiple experiments in less than four hours (Figure 7).
Figure 7. iWestern 4-hour workflow. By combining fast running Invitrogen Bolt Bis-Tris Plus gels, the rapid dry transfer of iBlot 3 Transfer Device, hands-free immunoprocessing of the iBind Flex western blot systems, and the easy, automatic workflows of iBright imaging systems, western blot experiments can be completed in less than 4 hours.
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Frequently asked questions
The iBind system performs membrane blocking, antibody incubation, and washing using sequential lateral flow to enable hands-free western blot processing.
No. The iBind system uses passive flow driven by capillary action and does not require external power or pumps.
Yes. Controlled reagent flow and reduced manual handling help improve consistency between experiments.
The iBind system helps provide a simple, hands-free workflow using passive flow, while the Bandmate system offers programmable control and higher throughput for more flexible workflows.
Choose the iBind system when simplicity, minimal setup, and ease of use are more important than customization or throughput.
Western blot educational resources
Access resources to help optimize western blot detection strategies, troubleshoot experiments, and improve data quality.
For Research Use Only. Not for use in diagnostic procedures.