How to perform a western blot semi-dry transfer using the Invitrogen Power Blotter and the Select Transfer Stacks


In this video, you’ll learn how to transfer proteins from polyacrylamide gels to membranes using the Invitrogen Power Blotter and Select Transfer Stacks. Invitrogen Power Blotter Select Transfer Stacks utilize a special application of the unique gel matrix technology developed for the Invitrogen iBlot 2 Transfer Stacks. Each stack contains the appropriate cathode and anode buffers in gel matrices, so there is no need to prepare transfer buffer. They also incorporate either PVDF or nitrocellulose membranes In the Power Blotter System, the cathode and anodes are plates that are fixed to the top and bottom of the cassette frame respectively, and the transfer stack is placed in between the cassette during the transfer process. The Power Blotter has two different cassette sizes: a standard size for transferring 1 midi gel or up to 2 mini gels, or the XL cassette for transferring up to 2 midi gels or up to 4 mini gels at a time. To begin the transfer process, flip the power switch at the rear of the device to turn on the system. Open the Power Blotter Cassette by pressing the grey button on the lid. Unseal the Select Transfer Stack and separate the top cathode stack. Set the top cathode stack to one side of the bench. Make sure the membrane isn’t stuck to the bottom of the separator. Remove the Bottom Anode Stack from the tray using forceps and place the Bottom Stack in the center of the anode. Use a roller to remove any bubbles from between the membrane and the transfer stack. Open the gel cassette and immerse the pre-run gel briefly in deionized water for 5 to 15 seconds. Gently shake off any excess water, then place the pre-run gel on the transfer membrane of the Bottom Anode Stack. Trim the gel carefully so that no parts of the gel hang over the sides of the stack. Use the roller to remove any air bubbles from between the gel and the membrane. Place the Top Cathode Stack, without the separator, on the pre-run gel and carefully roll to remove any air bubbles. Lock the top of the cassette, the cathode, into place and slide the assembled cassette into the Power Blotter Station. The Power Blotter system has preprogrammed transfer methods that are based on the number of gels being transferred and the molecular weight range of your proteins of interest. Custom methods can also be created. After selecting or creating a method, simply press Start to begin the transfer. The system signals the end of the transfer with repeated beeping sounds and a message on the display. Upon transfer completion, pull the cassette out from the base and open it. Carefully remove and discard the top stack and gel. Use forceps to remove the transfer membrane from the bottom stack. Membranes can rapidly dry out, so keep the membrane hydrated between each step in the procedure. Discard the bottom stack. When performing several consecutive transfers, drain the cassette by tilting it at the corner. Wipe down the anode surface with a damp cloth or paper tissue to remove any excess liquid. The Power Blotter system is ready for another run with no cooling period required unless the system has been running continuously for 2 hours. After extended use, cool the cassette for 30 minutes. Get more information about the Invitrogen Power Blotter at blotter

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