![]() ![]() A32723), Goat anti-Chicken Alexa Fluor 546 (Cat. MA3-414) followed by an incubation with secondary antibodies diluted at 1:5,000 in 1X blocking solution: Goat anti-Mouse Alexa Fluor Plus 488 (Cat. For fluorescence multiplex detection, the membranes were probed with a mixture of primary antibodies diluted in 1X blocking solution containing rabbit-anti EGFR (1:1,250) (Cat. Membranes were blocked in 1X Clear Milk Blocking Buffer. Membranes were processed using the Invitrogen Bandmate™ Automated Western Blot Processor. Proteins were transferred to either a 0.2 µm nitrocellulose membrane or a 0.45 µm PVDF membrane using the SureLock Tandem Midi Blot Module in the SureLock Tandem Midi Gel Tank at 25 V for 30 min. 10 µL of each A431 lysate concentration was loaded in each of 2 NuPAGE 4–12% Bis-Tris, 20-well gels and separated by electrophoresis using the SureLock Tandem Midi Gel Tank with the Invitrogen PowerEase Touch 350 W Power Supply and pre-programmed protocol NuPAGE BT (MOPS) (200 V 40 min). Multiplex western blotting following separation in the SureLock Tandem Midi Gel Tank and wet tank transfer using SureLock Tandem Midi Blot Modules generates excellent results on both nitrocellulose and PVDF membranes. A 2-fold dilution series of A431 cell lysate starting at 1.0 µg/µL was used along with iBright Prestained Protein Ladder. See the Ordering tab for more information.įigure 2. The SureLock Tandem Midi Gel Tank is also included with Invitrogen midi gels, buffers, and ladders, at no extra charge in several Protein Gels Welcome Packs. ![]() It can be purchased in a welcome pack with the SureLock Tandem Midi Blot Module and PVDF or nitrocellulose membranes, and SureLock Tandem Transfer Tray. The SureLock Tandem Midi Gel Tank can be purchased separately or in a variety of product bundles. The SureLock Tandem Midi Gel Tank is compatible with all Invitrogen precast midi gels and Invitrogen handcast midi gel cassettes, providing you with an effective and efficient 2-in-1 solution for producing publication-quality high throughput electrophoresis and western blotting results. Wet transfers can be performed in 30 minutes at room temperature, and there is no need to prechill buffers overnight or freeze cooling accessories before use. Separate chambers make it easy to run just one gel or transfer, allowing you to save on buffer and limit methanol waste. You can perform vertical electrophoresis of one or two midi gels, or transfer of one or two midi gels using the SureLock Tandem Midi Blot Module. The SureLock Tandem Midi Gel Tank is uniquely designed to enable convenient, reliable gel electrophoresis and protein transfer of high-performance Invitrogen midi gels. ![]()
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