Protein Electrophoresis and Western Blotting Education Center
Two devices, one easy technology for automated western blot processing
Invitrogen iBind Western Systems—no shakers, no trays, no timers. Set up in minutes and walk away. The Invitrogen iBind Western Systems are western blot processing benchtop devices that utilize sequential lateral flow (SLF) to perform hands-free blocking, antibody binding, and washes. In less than 3 hours, blots are processed and ready for final detection. You can continue to use your existing chromogenic, chemiluminescent, or fluorescent western blotting protocols, along with your choice of primary antibody or secondary antibody conjugates of HRP, AP, or fluorescent dyes.
What are the iBind Western Systems?
iBind Western Systems are small benchtop devices that automate the western blot processing (immunodetection) steps. The iBind Western Systems rely on a specialized glass fiber matrix, the iBind Card, to generate the sequential lateral flow (SLF) of immunodetection reagents for the blocking, antibody binding, and wash steps involved in the western blot immunodetection workflow.
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), i.e., simple capillary action—no electricity or batteries are required.
Figure 1. Time savings of automated western blotting processing with the iBind Western Systems.
Key benefits of the iBind Western Systems
- Small and portable—save bench space and easily store away in a drawer when not in use
- Easy set up—load reagents, assemble the prewetted iBind card with the membrane, close the lid and go. Your blot will be ready for detection in 3 hours.
- Flexible—compatible with PVDF and Nitrocellulose and all detection methods
- Reproducible results—consistent processing conditions help reduce blot-to-blot variations
Which iBind system is right for you?
Two iBind Western Systems are available: the original iBind Western Device, which accommodates the processing of one mini blot, and the iBind Flex Western Device, which accommodates the processing of up to two mini blots, one midi blot, or up to six vertically cut membrane strips at a time. Setup requires only approximately 15 minutes, with no additional hands-on steps until the blot has been processed and ready for the final detection steps.
| 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
Sequential lateral flow technology
The iBind Western Systems achieve automated blot processing without the use of 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 2 illustrates how this technology enables automated blot processing with the iBind Western Systems.
Click image to enlarge
Figure 2. Sequential lateral 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 System.
How to use iBind Western Systems
Learn how to process midi, mini or strip blots using the iBind Flex and process mini blots using the iBind with this detailed step by step video.
Results are as good as or better than manually processed western blots
- Automated processing enables improved blot-to-blot consistency with CVs typically less than 5% (compared to manual processing that can have CVs of 13%).
- Combine the iBind Western Systems with highly specific primary and secondary antibodies to achieve cleaner western blots.
Robust results with less primary antibody
Primary antibodies contribute to as much as 90% of the total cost of the blot. The iBind Western Systems require only 2 mL of primary antibody solution, helping to lower the cost per blot.
Figure 3. Using chemiluminescent or fluorescence-based detection methods, you can use up to 80% less primary antibody than manual methods, saving you significant cost per blot.
A 2-fold dilution series of EGF receptor control cell lysate (30 µg, 15 µg, 7.5 µg, 3.75 µg, and 1.875 µg) was used. Proteins were separated using Bolt Bis-Tris Plus gels on the Mini Gel Tank and transferred to PVDF membranes using the iBlot 2 Dry Blotting System. The blots were probed with an anti–phospho-EGF receptor [Tyr1068] (1H12) mouse monoclonal antibody (1:1,000 dilution, equated to 2 µL antibody for the iBind device method and 10 µL antibody for the manual method) followed by a goat anti–mouse IgG (H+L) peroxidase-conjugated antibody (1:360 for iBind device processing (5.55 µL); 1:1,800 for manual method (5.55 µL)). The standard iBind solution kit was used for the chemiluminescence blot (panel C, manual processing and panel D, iBind system processing); the fluorescence detection (panel A, manual processing and panel B, iBind processing) kit was used for fluorescence detection. These results demonstrate that western blots processed on the iBind device show comparable results to those obtained when western blots are processed manually, with lower overall primary antibody requirements for the iBind device.
Comparable results without the hands-on steps for midi, mini and strip blots
Figure 4. Comparison of manually processed midi blots vs. midi blots processed with the iBind Flex Western System.
Samples containing GST-tagged recombinant proteins were separated and then transferred to nitrocellulose membranes using the Invitrogen iBlot 2 Dry Blotting System. Blots were probed with identical concentrations of the same pair of primary and secondary antibodies. The primary antibody was rabbit anti-GST diluted 1:500 (8 µL in 4 mL iBind Flex Solution for the iBind system, 40 µL in 20 mL for manual tray incubation). The secondary antibody was goat anti-rabbit HRP diluted 1:600 (6.7 µL in 4 mL iBind Flex Solution for the iBind system, 33.3 µL in 20 mL for manual tray incubation). For final detection, blots were incubated for 5 minutes in Thermo Scientific SuperSignal West Dura Substrate for visualization with an imaging system.
Figure 5. Comparison of manually processed mini blots vs. mini blots processed with the iBind Flex Western System.
Blots were produced by separating samples and then transferred to nitrocellulose membranes using the iBlot 2 Dry Blotting System. Each gel contained 10 lanes loaded with two-fold dilution series of 293 cell extracts (30 µg to 0.06 µg). After immunodetection using the conditions described below, blots were incubated for 5 minutes in Thermo Scientific SuperSignal West Dura Substrate and the signal documented with an imager.
- MEK2 (45 kDa) – rabbit anti-MEK2 antibody applied at 1:1000 for iBind blot processing (= 2 µL in 2 mL of iBind Solution), and 1:1000 for manual blot processing (= 10 µL in 10 mL buffer).
- Securin(~28 kDa) – rabbit anti-Securin antibody applied at 1:1000 for iBind blot processing (= 2 µL in 2 mL of iBind Solution), and 1:1000 for manual blot processing (= 10 µL in 10 mL buffer).
- Secondary Antibody (both targets): goat anti-rabbit IgG at 1:600 for iBind blot processing (3.33 µL in 2 mL of iBind Solution), and at 1:1800 for manual blot processing (5.55 µL in 10 mL).
Figure 6. Comparison of 6 manually processed vertically cut blot strips vs. strips processed with the iBind Flex Western System.
Process up to 6 strips with 6 different antibodies combinations at a single time. Blots were produced by separating samples and transferring to nitrocellulose membranes using the iBlot 2 Dry Blotting System.
Get a jumpstart and save with our starter kits!
Our starter kits include everything you need to start processing your western blots using the iBind Western Systems.
iBind Western Device Starter kits
Option 1: iBind Western Starter Kit
- 1 iBind Western Device
- 10 iBind Cards, store at room temperature
- 1 iBind 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, store at room temperature
- 1 iBind Flex 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?
iBind Western Systems are small benchtop devices that automate the western blot processing (immunodetection) steps. The iBind Western Systems rely on a specialized glass fiber matrix, the iBind Card, to generate the sequential lateral flow (SLF) of immunodetection reagents for the blocking, antibody binding, and wash steps involved in the western blot immunodetection workflow.
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), i.e., simple capillary action—no electricity or batteries are required.
Figure 1. Time savings of automated western blotting processing with the iBind Western Systems.
Key benefits of the iBind Western Systems
- Small and portable—save bench space and easily store away in a drawer when not in use
- Easy set up—load reagents, assemble the prewetted iBind card with the membrane, close the lid and go. Your blot will be ready for detection in 3 hours.
- Flexible—compatible with PVDF and Nitrocellulose and all detection methods
- Reproducible results—consistent processing conditions help reduce blot-to-blot variations
Which iBind system is right for you?
Two iBind Western Systems are available: the original iBind Western Device, which accommodates the processing of one mini blot, and the iBind Flex Western Device, which accommodates the processing of up to two mini blots, one midi blot, or up to six vertically cut membrane strips at a time. Setup requires only approximately 15 minutes, with no additional hands-on steps until the blot has been processed and ready for the final detection steps.
| 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
Sequential lateral flow technology
The iBind Western Systems achieve automated blot processing without the use of 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 2 illustrates how this technology enables automated blot processing with the iBind Western Systems.
Click image to enlarge
Figure 2. Sequential lateral 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 System.
How to use iBind Western Systems
Learn how to process midi, mini or strip blots using the iBind Flex and process mini blots using the iBind with this detailed step by step video.
Results are as good as or better than manually processed western blots
- Automated processing enables improved blot-to-blot consistency with CVs typically less than 5% (compared to manual processing that can have CVs of 13%).
- Combine the iBind Western Systems with highly specific primary and secondary antibodies to achieve cleaner western blots.
Robust results with less primary antibody
Primary antibodies contribute to as much as 90% of the total cost of the blot. The iBind Western Systems require only 2 mL of primary antibody solution, helping to lower the cost per blot.
Figure 3. Using chemiluminescent or fluorescence-based detection methods, you can use up to 80% less primary antibody than manual methods, saving you significant cost per blot.
A 2-fold dilution series of EGF receptor control cell lysate (30 µg, 15 µg, 7.5 µg, 3.75 µg, and 1.875 µg) was used. Proteins were separated using Bolt Bis-Tris Plus gels on the Mini Gel Tank and transferred to PVDF membranes using the iBlot 2 Dry Blotting System. The blots were probed with an anti–phospho-EGF receptor [Tyr1068] (1H12) mouse monoclonal antibody (1:1,000 dilution, equated to 2 µL antibody for the iBind device method and 10 µL antibody for the manual method) followed by a goat anti–mouse IgG (H+L) peroxidase-conjugated antibody (1:360 for iBind device processing (5.55 µL); 1:1,800 for manual method (5.55 µL)). The standard iBind solution kit was used for the chemiluminescence blot (panel C, manual processing and panel D, iBind system processing); the fluorescence detection (panel A, manual processing and panel B, iBind processing) kit was used for fluorescence detection. These results demonstrate that western blots processed on the iBind device show comparable results to those obtained when western blots are processed manually, with lower overall primary antibody requirements for the iBind device.
Comparable results without the hands-on steps for midi, mini and strip blots
Figure 4. Comparison of manually processed midi blots vs. midi blots processed with the iBind Flex Western System.
Samples containing GST-tagged recombinant proteins were separated and then transferred to nitrocellulose membranes using the Invitrogen iBlot 2 Dry Blotting System. Blots were probed with identical concentrations of the same pair of primary and secondary antibodies. The primary antibody was rabbit anti-GST diluted 1:500 (8 µL in 4 mL iBind Flex Solution for the iBind system, 40 µL in 20 mL for manual tray incubation). The secondary antibody was goat anti-rabbit HRP diluted 1:600 (6.7 µL in 4 mL iBind Flex Solution for the iBind system, 33.3 µL in 20 mL for manual tray incubation). For final detection, blots were incubated for 5 minutes in Thermo Scientific SuperSignal West Dura Substrate for visualization with an imaging system.
Figure 5. Comparison of manually processed mini blots vs. mini blots processed with the iBind Flex Western System.
Blots were produced by separating samples and then transferred to nitrocellulose membranes using the iBlot 2 Dry Blotting System. Each gel contained 10 lanes loaded with two-fold dilution series of 293 cell extracts (30 µg to 0.06 µg). After immunodetection using the conditions described below, blots were incubated for 5 minutes in Thermo Scientific SuperSignal West Dura Substrate and the signal documented with an imager.
- MEK2 (45 kDa) – rabbit anti-MEK2 antibody applied at 1:1000 for iBind blot processing (= 2 µL in 2 mL of iBind Solution), and 1:1000 for manual blot processing (= 10 µL in 10 mL buffer).
- Securin(~28 kDa) – rabbit anti-Securin antibody applied at 1:1000 for iBind blot processing (= 2 µL in 2 mL of iBind Solution), and 1:1000 for manual blot processing (= 10 µL in 10 mL buffer).
- Secondary Antibody (both targets): goat anti-rabbit IgG at 1:600 for iBind blot processing (3.33 µL in 2 mL of iBind Solution), and at 1:1800 for manual blot processing (5.55 µL in 10 mL).
Figure 6. Comparison of 6 manually processed vertically cut blot strips vs. strips processed with the iBind Flex Western System.
Process up to 6 strips with 6 different antibodies combinations at a single time. Blots were produced by separating samples and transferring to nitrocellulose membranes using the iBlot 2 Dry Blotting System.
Get a jumpstart and save with our starter kits!
Our starter kits include everything you need to start processing your western blots using the iBind Western Systems.
iBind Western Device Starter kits
Option 1: iBind Western Starter Kit
- 1 iBind Western Device
- 10 iBind Cards, store at room temperature
- 1 iBind 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, store at room temperature
- 1 iBind Flex 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
See what our customers are saying about iBind
iBind Western Device is easy to use
iBind Western Device saves time and is consistent
Use less primary antibody with iBind Western Device
Resources
Download: The iWestern Workflow Brochure
Download: Western Detection Workflow Brochure
Explore: Western Blot Validated Antibodies
For Research Use Only. Not for use in diagnostic procedures.