The iBind™ Western System is an automated western blot processing platform that requires less primary antibody and delivers sensitive, reproducible western results. All blocking, antibody incubation, and washing steps are hands-free, allowing you to load your solutions and walk away. There is no electricity or battery required. You can also use your existing chemiluminescent, chromogenic, or fluorescent western detection protocols, including HRP-, AP-, Alexa Fluor® dye–, and IRDye® (LI-COR®)–conjugated secondary antibodies.
- Cost savings—use up to 80% less primary antibody*
- Superior sensitivity—detect proteins at lower levels than manually processed blots
- Reproducibility—automated processing enables improved blot-to-blot consistency
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.
Figure 1. 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 extract (30 µg, 15 µg, 7.5 µg, 3.75 µg, and 1.875 µg). Proteins were separated using the Mini Gel Tank Electrophoresis System and transferred to PVDF membranes using the iBlot® 7-Minute Blotting System. The blots were probed with a 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™ Western System offers greater sensitivity compared to manual methods for many monoclonal and polyclonal antibodies. Figures 1 and 2 compare the level of sensitivity for the iBind™ System versus manual western methods.
- Combine the iBind™ Western System with highly specific Novex® primary and secondary antibodies to achieve cleaner western blots.
- Automated processing provides improved blot-to-blot consistency with CVs typically less than 5% (compared to manual processing that can have CVs of 13%).
||Figure 2. Western blots processed on the iBind™ device show superior sensitivity compared to western blots processed manually. (A–B) Western blots with phosphorylated Akt (left to right: 30 µg to 500 ng cell extract) were processed either on the iBind™ device or using standard manual western processing protocols as specified by the antibody manufacturer (monoclonal anti–phospho-Akt[Thr308] (C31E5E) primary antibody; HRP-conjugated anti-rabbit secondary antibody). The blot processed with the iBind™ device detected phospho-Akt in 500 ng of cell extract, while the target was detected at 4 µg on the manually processed blot. (C, D) Western blots with cell lysate expressing CREB (left to right: 30 µg to 1 µg cell lysate) 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 the Bolt® Gel Electrophoresis System and transferred to PVDF membranes using the iBlot® 7-Minute Blotting System.
|Figure 3. 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 to 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 [Tyr705] (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 Bolt® Gel Electrophoresis System and transferred to nitrocellulose membranes using the iBlot® 2 Dry Blotting System.
Get your iBind™ Western System Starter Kit now for only $999
Take advantage of faster, more sensitive and consistent western blots. In addition to the iBind™ Western System, the iBind™ Starter Kit provides cards and a solution kit, so you can get started right away. With the starter kit, you'll get:
- iBind™ Western System
- 1 box of iBind™ Cards
- iBind™ Solution Kit*
* Use the iBind™ Fluorescent Detection (FD) Solution Kit for infrared fluorescent-based detection with LI-COR® ODYSSEY® imagers.
Order the iBind™ western system starter kit now
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