one pre-wetted fiber pad on the gray side of the cassette. Place a sheet of filter paper on the fiber pad. Place the equilibrated gel on the filter paper. Remove any air bubbles under the gel or the pre-wetted membrane membrane by rolling them out. Use a on the gel. specialized roller or simply a glass test tube. Place a piece of filter paper on the membrane. Place a fiber pad on the filter paper. Close the cassette firmly, being careful not to move the gel and filter paper sandwich. Lock the cassette closed with the latch. Place the assembled cassette into the module. Place the module into the tank with the black electrode to the top and left. Add a cooling unit to the tank. Fill the tank with buffer to the mark “blotting” on the tank. Add a stir bar to the tank and Stirring the buffer will help maintain a cool place the tank on top of a stir plate. Set and even buffer temperature and the stirring speed as fast as possible homogeneous ion concentration. without splashing. The lid only fits on one way, so make sure the lid on the tank, matching the the module is oriented correctly. electrode colors. PART B: RUNNING THE GEL The voltage, current, and time should be Connect the electrical leads to a selected based on the type of buffer, gels, power supply. Set to an appropriate and analyte(s). One option for SDS-PAGE voltage and current and run the gels containing proteins in TG buffer with transfer. alcohol would be a one-hour transfer at 100 V and 250 mA. After starting, check the area around the electrodes to make sure tiny bubbles are forming. This is evidence of an electrical current in the buffer.
one pre-wetted fiber pad on the gray side of the cassette. Place a sheet of filter paper on the fiber pad. Place the equilibrated gel on the filter paper. the pre-wetted membrane on the gel. Place a piece of filter paper on the membrane. Place a fiber pad on the filter paper. Close the cassette firmly, being careful not to move the gel and filter paper sandwich. Lock the cassette closed with the latch. Place the assembled cassette into the module. Place the module into the tank with the black electrode to the top and left. Add a cooling unit to the tank. Fill the tank with buffer to the “blotting” mark on the tank. Add a stir bar to the tank and place the tank on top of a stir plate. Set the stirring speed as fast as possible without splashing. the lid on the tank, matching the electrode colors. PART B: RUNNING THE GEL Connect the electrical leads to a power supply. Set to an appropriate voltage and current and run the transfer. After starting, check the area around the electrodes to make sure tiny bubbles are forming. This is evidence of an electrical current in the buffer. Remove any air bubbles under the gel or membrane by rolling them out. Use a specialized roller or simply a glass test tube. Stirring the buffer will help maintain a cool and even buffer temperature and homogeneous ion concentration. The lid only fits on one way, so make sure the module is oriented correctly. The voltage, current, and time should be selected based on the type of buffer, gels, and analyte(s). One option for SDS-PAGE gels containing proteins in TG buffer with alcohol would be a one-hour transfer at 100 V and 250 mA.
2012_fall_sop_book_9th_ed_1.docx