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iBind Automated Western Systems
Two devices, one easy technology for automated western blot processing
Load and go—set up in minutes, walk away and return to detection-ready blots in under 3 hours. Automate your workflow with Invitrogen iBind Western Systems and eliminate tedious blocking, antibody binding and washing steps, while reducing variability and improving sensitivity. Your antibodies, our technology—easily adapt your protocol to the iBind sequential lateral flow technology—no shaker, trays or timers needed.
What are the iBind Western Systems?
The iBind Western Systems consist of the iBind Western Device, iBind sequential lateral flow cards, and iBind solution kits (available for chemiluminescent or fluorescent detection). The iBind Western Systems are available with two throughput options—iBind Western System for one mini blot or iBind Flex Western System for two mini blots, one midi blot or vertically cut strips.
Streamlined workflow—load and go
iBind Western Systems allow all solutions to be prepared and loaded in the device at the start of the procedure, from which all subsequent steps proceed automatically and uninterrupted by sequential lateral flow technology (SLF), e.g., simple capillary action—no electricity or batteries are required. 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 System.
Additional benefits of the iBind Western Systems
- Reproducible results—consistent processing conditions help reduce blot-to-blot variations.
- Reagent cost efficiency—the iBind PLUS Solution Kit lowers antibody volume requirements and helps reduce costs compared with manual processing.
- Flexible—compatible with PVDF and Nitrocelluose and all detection methods.
- Small and portable—save bench space and easily store away in a drawer when not in use.
Figure 2. Use 5X less 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 (Cat. No. IB33002) using the iBlot 3 Transfer Device (Cat. No. IB31001). Membranes were then incubated with primary and secondary antibodies for the detection of 3 different protein targets (4EBP1, Caspase 7, AKT) using either iBind Western System with iBind PLUS Solution Kit or standard western processing of manual blocking, washing and antibody incubation. Primary antibody concentration of 1:1000 was used for all western blots, while higher concentration of secondary antibody was used for iBind Western Systems blot processing (1:10,000) vs manual western blot processing (1:100,000).
Which iBind system is right for you?
Two iBind Western Systems are available: the original iBind Western Device, and the iBind Flex Western Device. Setup requires only approximately 15 minutes, with no additional hands-on steps until the blot has been processed and is ready for the final detection steps.
Reference the table below to choose the device that meets the needs of your lab.
| iBind Western Device | iBind Flex Western Device | |
|---|---|---|
 |  | |
| Mini blot (single) | Yes | Yes |
| Mini blot (dual) | No | Yes |
| Midi blot | No | Yes |
| Vertically cut strips | No | Yes |
Flexible 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.
Midi blots
Using the same antibody conditions
Mini blots
Using the same or different antibody conditions
Vertically cut strips
Using the same or different antibody conditions
iBind Cards and iBind Flex Cards and solution kits
iBind Cards 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.
iBind and iBind Flex solution kits
iBind solution kits are available for either chemiluminescent or fluorescent detection. The iBind PLUS Solution Kits and iBind Flex PLUS for chemiluminescent detection contain ready-to-use reagents including a buffer solution for blocking, wetting and wash steps and a diluent for primary and secondary antibody preparation. The iBind Fluorescent Detection (FD) Solution Kit and iBind Flex Fluorescent Detection (FD) Solution Kit contain concentrated buffer and additive that, when prepared, are used for blocking, wash steps, and antibody dilution. A 10% SDS detergent is supplied in both the iBind FD Solution Kit and iBind Flex FD Solution Kit for addition to fluorescently-conjugated secondary antibodies. iBind FD Solution Kit and iBind Flex FD Solution Kit enable fast multiplex fluorescent detection and low background when used with iBind Western Systems.
Sequential lateral flow technology
The iBind Western Systems achieve automated blot processing without the use of a complex, high-maintenance fluidics system. Instead, they rely on sequential lateral flow technology (SLF). Similar to rapid diagnostic tests, SFL is based on the ability of a liquid to move through paper or a specialized matrix via capillary action. Figure 3 illustrates how this technology enables automated blot processing with the iBind Western Systems.
Figure 3. Sequential lateral flow technology employed by the iBind Western Systems.
Sequential lateral flow technology explained
Learn how the application of sequential lateral flow technology allows automated western blot processing with the iBind Western Device.
How to use iBind Western Systems
Learn how to process midi, mini or strip blots using the iBind Flex Western Device and process mini blots using the iBind Western Device with this detailed step by step video.
Results with iBind automated western systems are as good or better than manually processed western blots
The iBind systems and sequential lateral flow offers more than hands-free automation and efficiency. It can also improve your western blot results, help save on antibody costs, and simplify experiments examining multiple protein targets.
Sequential lateral flow moves the antibody and wash solutions directly and evenly across the membrane. This results in minimal diffusion requirements for target binding and unbound antibody is quickly washed away without the need for multiple wash cycles. This reduces volume of reagents and antibodies required and allows for high concentrations of secondary antibodies to be used to further enhance detection without high background.
High sensitivity chemiluminescent detection with less primary antibody using iBind PLUS and iBind Flex PLUS solution kits
Primary antibodies can contribute to as much as 60–80% of the total cost of a western blot experiment. Standard antibody processing of western blots is typically conducted with 10 mL of a primary antibody at dilution ranges typically between 1:1,000 to 1:5,000 in order to properly cover the western blot membrane and enable enough binding of primary antibody to the target protein. In contrast, iBind Western Systems uses only 2 mL of diluted primary antibody, often at the same or similar concentration to manual processing protocols. This represents an 80% reduction in the amount of primary antibody used.
In addition to the reduced antibody volume required, the antibody diluent in the iBind PLUS Solution Kit and iBind Flex PLUS Solution Kit are specially formulated to increase and enhance antibody binding kinetics between the primary antibody and target protein. When using the iBind PLUS Solution Kit, increased signal is often obtained compared to standard overnight western blots when using the same antibody dilutions (Figure 4). This means that for many protein targets, primary antibody usage can be further reduced to generate similar detection sensitivity as traditional methods (Figure 5).
Figure 4. iBind Western Systems with iBind PLUS Solution kits demonstrate consistent and comparable 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 (Cat. No. IB33002) using the iBlot 3 Transfer Device (Cat. No. IB31001). Membranes were then incubated with primary and secondary antibodies using either iBind Western Systems and iBind PLUS Solution Kit (left side panel) or by manual processing (right side panel) 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 (Cat. No. XP04202BOX), and transferred on NC membrane using the iBlot 3 Western Blot Transfer System (Cat. No. IB31001 and IB33002). Membranes were then processed with primary antibodies for 3 targets (AKT, Ezrin, and MEK1) using AKT Pan Monoclonal Antibody (Cat. No. MA5-14916), Ezrin Monoclonal Antibody (Cat. No. 35-7300), or MEK1 Monoclonal Antibody (Cat. No. 13-3500), respectively, at varying concentrations as indicated using the iBind Western Systems with iBind PLUS Solution Kit, followed by incubation with secondary-HRP antibody diluted at 1:10,000 as recommended for the iBind Western Systems protocol. Manual western blotting was processed using Pierce Clear Milk Blocking Buffer (Cat. No. 37587) 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 (~17–18 hours). 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.
Increase productivity using the iBind Flex device as part of the iWestern workflow
The iBind Flex Western Device is specifically designed to support flexible membrane formats and enable the ability to run larger experiments. The iBind Flex system comes with interchangeable wells, which allows you to run multiple membrane formats and/or run different primary and secondary antibody conditions on the same device at the same time. The iBind Flex device supports the processing of up to two mini blots, one midi blot, or up to six vertically cut membrane strips at a time.
When combining the iBind Flex device and the iBind Flex PLUS kit (for chemiluminescent detection) or iBind Flex FD kit (for fluorescent detection), researchers can significantly improve productivity compared to overnight incubation. When paired with with the broader iWestern workflow system of fast, precast Invitrogen Bolt Bis-Tris protein gels, modern Mini Gel Tank and Power Ease Touch power supplies, rapid iBlot 3 dry transfer system and the iBright FL1500 imager, researchers can complete single to multiple multiplexed experiments in less than four hours (Figure 5 and Figure 6).
Figure 6. iWestern workflow. By combining fast running Invitrogen Bolt Bis-Tris Plus gels, the rapid dry transfer of iBlot 3 transfer system, 3-hour hands-free immunoprocessing of the iBind Flex Western Systems, and the easy, automatic workflows of iBright gel and western blot imaging systems, western blot experiments can be completed in less than 4 hours.
Figure 7. Similar or greater detection sensitivity in multiplex experiments. NuPAGE Bis-Tris 4–12% precast gels (Cat. No. NP0322BOX) were loaded with dilution series (2-Fold) of cell lysate and then separated and transferred to nitrocellulose membranes (Cat. No. IB33002) using the iBlot 3 Transfer Device (Cat. No. IB31001). Membranes were then incubated with primary and secondary antibodies using either iBind Flex Western 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 System 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 antibody (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.
Get a jumpstart with our starter kits
Our starter kits include everything you need to start processing your western blots using the iBind Western Systems.
iBind Western Starter kits
Option 1: iBind Western Starter Kit
- 1 iBind Western Device
- 10 iBind Cards
- 1 iBind PLUS Solution Kit
Option 2: iBind Western Starter Kit, Fluorescent Detection
- 1 iBind Western Device
- 10 iBind Cards, store at room temperature
- 1 iBind Fluorescent Detection (FD) Solution Kit
iBind Flex Western Device Starter kits
Option 1: iBind Flex Western Starter Kit
- 1 iBind Flex Western Device
- 10 iBind Flex Cards
- 1 iBind Flex PLUS Solution Kit
Option 2: iBind Flex Western Starter Kit, Fluorescent Detection
- 1 iBind Flex Western Device
- 10 iBind Flex Cards, store at room temperature
- 1 iBind Flex Fluorescent Detection (FD) Solution Kit
Manuals
What are the iBind Western Systems?
The iBind Western Systems consist of the iBind Western Device, iBind sequential lateral flow cards, and iBind solution kits (available for chemiluminescent or fluorescent detection). The iBind Western Systems are available with two throughput options—iBind Western System for one mini blot or iBind Flex Western System for two mini blots, one midi blot or vertically cut strips.
Streamlined workflow—load and go
iBind Western Systems allow all solutions to be prepared and loaded in the device at the start of the procedure, from which all subsequent steps proceed automatically and uninterrupted by sequential lateral flow technology (SLF), e.g., simple capillary action—no electricity or batteries are required. 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 System.
Additional benefits of the iBind Western Systems
- Reproducible results—consistent processing conditions help reduce blot-to-blot variations.
- Reagent cost efficiency—the iBind PLUS Solution Kit lowers antibody volume requirements and helps reduce costs compared with manual processing.
- Flexible—compatible with PVDF and Nitrocelluose and all detection methods.
- Small and portable—save bench space and easily store away in a drawer when not in use.
Figure 2. Use 5X less 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 (Cat. No. IB33002) using the iBlot 3 Transfer Device (Cat. No. IB31001). Membranes were then incubated with primary and secondary antibodies for the detection of 3 different protein targets (4EBP1, Caspase 7, AKT) using either iBind Western System with iBind PLUS Solution Kit or standard western processing of manual blocking, washing and antibody incubation. Primary antibody concentration of 1:1000 was used for all western blots, while higher concentration of secondary antibody was used for iBind Western Systems blot processing (1:10,000) vs manual western blot processing (1:100,000).
Which iBind system is right for you?
Two iBind Western Systems are available: the original iBind Western Device, and the iBind Flex Western Device. Setup requires only approximately 15 minutes, with no additional hands-on steps until the blot has been processed and is ready for the final detection steps.
Reference the table below to choose the device that meets the needs of your lab.
| iBind Western Device | iBind Flex Western Device | |
|---|---|---|
| | |
| Mini blot (single) | Yes | Yes |
| Mini blot (dual) | No | Yes |
| Midi blot | No | Yes |
| Vertically cut strips | No | Yes |
Flexible 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.
Midi blots
Using the same antibody conditions
Mini blots
Using the same or different antibody conditions
Vertically cut strips
Using the same or different antibody conditions
iBind Cards and iBind Flex Cards and solution kits
iBind Cards 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.
iBind and iBind Flex solution kits
iBind solution kits are available for either chemiluminescent or fluorescent detection. The iBind PLUS Solution Kits and iBind Flex PLUS for chemiluminescent detection contain ready-to-use reagents including a buffer solution for blocking, wetting and wash steps and a diluent for primary and secondary antibody preparation. The iBind Fluorescent Detection (FD) Solution Kit and iBind Flex Fluorescent Detection (FD) Solution Kit contain concentrated buffer and additive that, when prepared, are used for blocking, wash steps, and antibody dilution. A 10% SDS detergent is supplied in both the iBind FD Solution Kit and iBind Flex FD Solution Kit for addition to fluorescently-conjugated secondary antibodies. iBind FD Solution Kit and iBind Flex FD Solution Kit enable fast multiplex fluorescent detection and low background when used with iBind Western Systems.
Sequential lateral flow technology
The iBind Western Systems achieve automated blot processing without the use of a complex, high-maintenance fluidics system. Instead, they rely on sequential lateral flow technology (SLF). Similar to rapid diagnostic tests, SFL is based on the ability of a liquid to move through paper or a specialized matrix via capillary action. Figure 3 illustrates how this technology enables automated blot processing with the iBind Western Systems.
Figure 3. Sequential lateral flow technology employed by the iBind Western Systems.
Sequential lateral flow technology explained
Learn how the application of sequential lateral flow technology allows automated western blot processing with the iBind Western Device.
How to use iBind Western Systems
Learn how to process midi, mini or strip blots using the iBind Flex Western Device and process mini blots using the iBind Western Device with this detailed step by step video.
Results with iBind automated western systems are as good or better than manually processed western blots
The iBind systems and sequential lateral flow offers more than hands-free automation and efficiency. It can also improve your western blot results, help save on antibody costs, and simplify experiments examining multiple protein targets.
Sequential lateral flow moves the antibody and wash solutions directly and evenly across the membrane. This results in minimal diffusion requirements for target binding and unbound antibody is quickly washed away without the need for multiple wash cycles. This reduces volume of reagents and antibodies required and allows for high concentrations of secondary antibodies to be used to further enhance detection without high background.
High sensitivity chemiluminescent detection with less primary antibody using iBind PLUS and iBind Flex PLUS solution kits
Primary antibodies can contribute to as much as 60–80% of the total cost of a western blot experiment. Standard antibody processing of western blots is typically conducted with 10 mL of a primary antibody at dilution ranges typically between 1:1,000 to 1:5,000 in order to properly cover the western blot membrane and enable enough binding of primary antibody to the target protein. In contrast, iBind Western Systems uses only 2 mL of diluted primary antibody, often at the same or similar concentration to manual processing protocols. This represents an 80% reduction in the amount of primary antibody used.
In addition to the reduced antibody volume required, the antibody diluent in the iBind PLUS Solution Kit and iBind Flex PLUS Solution Kit are specially formulated to increase and enhance antibody binding kinetics between the primary antibody and target protein. When using the iBind PLUS Solution Kit, increased signal is often obtained compared to standard overnight western blots when using the same antibody dilutions (Figure 4). This means that for many protein targets, primary antibody usage can be further reduced to generate similar detection sensitivity as traditional methods (Figure 5).
Figure 4. iBind Western Systems with iBind PLUS Solution kits demonstrate consistent and comparable 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 (Cat. No. IB33002) using the iBlot 3 Transfer Device (Cat. No. IB31001). Membranes were then incubated with primary and secondary antibodies using either iBind Western Systems and iBind PLUS Solution Kit (left side panel) or by manual processing (right side panel) 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 (Cat. No. XP04202BOX), and transferred on NC membrane using the iBlot 3 Western Blot Transfer System (Cat. No. IB31001 and IB33002). Membranes were then processed with primary antibodies for 3 targets (AKT, Ezrin, and MEK1) using AKT Pan Monoclonal Antibody (Cat. No. MA5-14916), Ezrin Monoclonal Antibody (Cat. No. 35-7300), or MEK1 Monoclonal Antibody (Cat. No. 13-3500), respectively, at varying concentrations as indicated using the iBind Western Systems with iBind PLUS Solution Kit, followed by incubation with secondary-HRP antibody diluted at 1:10,000 as recommended for the iBind Western Systems protocol. Manual western blotting was processed using Pierce Clear Milk Blocking Buffer (Cat. No. 37587) 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 (~17–18 hours). 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.
Increase productivity using the iBind Flex device as part of the iWestern workflow
The iBind Flex Western Device is specifically designed to support flexible membrane formats and enable the ability to run larger experiments. The iBind Flex system comes with interchangeable wells, which allows you to run multiple membrane formats and/or run different primary and secondary antibody conditions on the same device at the same time. The iBind Flex device supports the processing of up to two mini blots, one midi blot, or up to six vertically cut membrane strips at a time.
When combining the iBind Flex device and the iBind Flex PLUS kit (for chemiluminescent detection) or iBind Flex FD kit (for fluorescent detection), researchers can significantly improve productivity compared to overnight incubation. When paired with with the broader iWestern workflow system of fast, precast Invitrogen Bolt Bis-Tris protein gels, modern Mini Gel Tank and Power Ease Touch power supplies, rapid iBlot 3 dry transfer system and the iBright FL1500 imager, researchers can complete single to multiple multiplexed experiments in less than four hours (Figure 5 and Figure 6).
Figure 6. iWestern workflow. By combining fast running Invitrogen Bolt Bis-Tris Plus gels, the rapid dry transfer of iBlot 3 transfer system, 3-hour hands-free immunoprocessing of the iBind Flex Western Systems, and the easy, automatic workflows of iBright gel and western blot imaging systems, western blot experiments can be completed in less than 4 hours.
Figure 7. Similar or greater detection sensitivity in multiplex experiments. NuPAGE Bis-Tris 4–12% precast gels (Cat. No. NP0322BOX) were loaded with dilution series (2-Fold) of cell lysate and then separated and transferred to nitrocellulose membranes (Cat. No. IB33002) using the iBlot 3 Transfer Device (Cat. No. IB31001). Membranes were then incubated with primary and secondary antibodies using either iBind Flex Western 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 System 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 antibody (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.
Get a jumpstart with our starter kits
Our starter kits include everything you need to start processing your western blots using the iBind Western Systems.
iBind Western Starter kits
Option 1: iBind Western Starter Kit
- 1 iBind Western Device
- 10 iBind Cards
- 1 iBind PLUS Solution Kit
Option 2: iBind Western Starter Kit, Fluorescent Detection
- 1 iBind Western Device
- 10 iBind Cards, store at room temperature
- 1 iBind Fluorescent Detection (FD) Solution Kit
iBind Flex Western Device Starter kits
Option 1: iBind Flex Western Starter Kit
- 1 iBind Flex Western Device
- 10 iBind Flex Cards
- 1 iBind Flex PLUS Solution Kit
Option 2: iBind Flex Western Starter Kit, Fluorescent Detection
- 1 iBind Flex Western Device
- 10 iBind Flex Cards, store at room temperature
- 1 iBind Flex Fluorescent Detection (FD) Solution Kit
Manuals
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iBind Western Device is easy to use
iBind Western Device helps saves time and is consistent
Use less primary antibody with iBind Western Device
Stylesheet for Classic Wide Template adjustments
For Research Use Only. Not for use in diagnostic procedures.