The Invitrogen™ iBind™ Western System for mini blots is an automated western blot processing platform that requires less primary antibody and enables sensitive, reproducible western results. All blocking, antibody incubation, and washing steps are hands-free, allowing you to load your solutions and walk away. No electricity or battery is required. You can also use your existing chemiluminescent, chromogenic, or fluorescent western detection protocols, including primary or secondary antibody conjugates of HRP, AP, or fluorescent dyes.

  • Cost savings—use up to 80% less primary antibody than with traditional tray-based incubation steps for western blotting
  • Reproducibility—automated processing enables improved blot-to-blot consistency
  • Simplicity—the system processes solutions using passive sequential lateral flow technology; no batteries or electricity required

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Example data

Robust results with less primary antibody

One of the key elements of a successful western blot is the primary antibody; however, this reagent also contributes to over 90% of the total cost of the blot. Because the iBind™ Western System is more sensitive than manual processing methods, you can use less antibody and achieve similar results.

*Results may vary. The protocol for primary antibody use is 80% less than a traditional manual method.

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.

Superior western performance compared to manual blotting
  • The iBind™ System offers greater sensitivity compared to manual methods for many monoclonal and polyclonal antibodies.
  • Combine the iBind™ Western System with highly specific primary and secondary antibodies to achieve cleaner 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%).
  Western blots processed on the Invitrogen™ iBind™ device show superior sensitivity compared to western blots processed manually. (A–B) Western blots with phosphorylated Akt (left to right: 30 µg–500 ng cell lysate load) were processed either on the iBind device or using standard manual western processing protocols as specified by the antibody manufacturer (monoclonal anti–phospho-Akt [pT308] (C31E5E) primary antibody; HRP-conjugated anti-rabbit secondary antibody). The blot processed with the iBind device detected phospho-Akt in 500 ng of cell lysate, while the target was detected in 4 µg on the manually processed blot. (C–D) Western blots with cell lysate expressing CREB (left to right: 30 µg–1 µg cell lysate load) were processed either on the iBind device or using standard manual western processing protocols as specified by the antibody manufacturer (polyclonal anti-CREB primary antibody; HRP-conjugated anti-rabbit secondary antibody). The blot processed with the iBind system detected CREB in 6 µg of cell lysate, while 10 µg of lysate was needed to detect CREB on the manually processed blot. For all blots, 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.

Invitrogen™ iBind™ system vs. manual western blotting with fluorescence-based detection of c-Jun and phospho-Stat3. Western blots of cell lysates containing phospho-Stat3 and c-Jun (left to right: 30 µg–120 ng lysate protein) were processed either on the iBind device or using standard manual western processing protocols as specified by the manufacturer for each antibody (monoclonal phospho-Stat3 [pY705] (M9C6) mouse primary antibody and monoclonal c-Jun (60A8) rabbit primary antibody). (A), (C) Alexa Fluor™ 680 goat anti-rabbit secondary antibody and Alexa Fluor™ 790 goat anti-mouse secondary antibody. (B), (D) IRDye™ 680LT goat anti-rabbit secondary antibody and IRDye™ 800CW goat anti-mouse secondary antibody. Blots processed with the iBind device detected both target proteins at lower levels than manually processed blots. For all blots, proteins were separated using the Invitrogen™ Bolt™ Gel Electrophoresis System and transferred to NC membranes using the Invitrogen™ iBlot™ 2 Dry Blotting System.

Get your iBind™ Western System Starter Kit now

Begin using a simpler, more elegant western blotting process. The iBind Starter Kit includes a complete iBind Western Device, plus cards and a solution kit, so you can get started right away:

  • iBind™ Western System
  • iBind Cards (1 box of 10)
  • iBind Solution Kit (Note: fluorescence detection requires FD Solution Kit)

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Everything you need for high-quality separation
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Sample of available secondary antibodies