How to perform a western blot dry transfer using the Invitrogen iBlot 2 Dry Blotting System

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In this video, you’ll learn how to transfer proteins from polyacrylamide gels to membranes using the Invitrogen iBlot2 Dry Blotting System. The iBlot2 Dry Blotting System uses pre-assembled transfer stacks with integrated nitrocellulose or PVDF membranes, offering the convenience of a bufferless plug and play system. iBlot 2 stack consists of copper cathode and anode sandwiching dry gel matrices containing the appropriate cathode and anode buffers so there is no need to prepare transfer buffer. Both mini and midi blot stacks are available—mini blot stacks transfer 1 mini blot, while regular blot stacks can transfer 1 midi blot or up to 2 mini blots. To get started, turn on the iBlot 2 dry transfer device and open the lid. The digital display will show icons for the available actions. Next, unseal the transfer stack. Notice that the top and bottom stacks are in the same tray. Separate the top stack and set it aside. Keep the bottom stack in the plastic tray. The bottom stack contains your PVDF or nitrocellulose membrane. Place the tray on the blotting surface of the iBlot2. Be sure to align the electrical contacts on the tray with the corresponding contacts on the blotting surface. Next, pick up your pre-run gel and place it on the membrane. Pre-soak an iBlot filter paper with deionized water and place it on top of the gel. Then remove air bubbles using the roller. Place the top stack over the pre-soaked filter paper and remove air bubbles using the roller. Place the absorbent pad on top of the top stack. Again, be sure that electrical contacts are aligned with the corresponding contacts on the blotting surface. Then close the lid. You have the ability to create custom methods of your own, or choose from our recommended preset templates. Select the desired method and make sure that the parameters are correct. Touch START to run, or use the “start last run” icon. Once the transfer begins, the elapsed time is displayed on the screen. The end of the transfer is indicated by beeping sounds and a message on the digital display. To disassemble, open the lid of the device, carefully remove and discard the top stack and the gel. Remove the transfer membrane from the stack and either place it in water or blocking solution. The bottom stack can now be discarded. Your membrane is ready for western detection. To clean the iBlot2 system, make sure the system is turned off, then wipe off the blotting surface and electrodes with a damp cloth or tissue paper. Get more information about the Invitrogen iBlot 2 Dry Blotting system at thermofisher.com/iblot2

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