DYCP-40C semi-dry blotting system is used together with electrophoresis power supply for transferring the proteins in the polyacrylamide gels onto the membrane like nitrocellulose membrane, nylon membrane and PVDF membrane. Semi-dry blotting is performed with graphite plate electrodes in a horizontal configuration, sandwiching a gel and membrane between sheets of buffer-soaked filter paper that function as the ion reservoir. During the electrophoretic transfer, negatively charged molecules migrate out of the gel and move towards the positive electrode, where they are deposited on the membrane. The plate electrodes, separated only by the gel and filter paper stack, provide high field strength (V/cm) across the gel, carrying out the very efficient, rapid transfers. The transfer surface of the smaller DYCP – 40C electrophoresis cell is 150 × 150 (mm), suitable to transferring standard gels, including those from DYCZ-24DN and DYCZ-24EN electrophoresis cell.
Let us learn more about the operation of this semi-dry trans blot apparatus.
Materials, instruments for operating DYCP-40C
Electrophoresis power supply DYY-6C, semi-dry trans blot apparatus DYCP-40C, buffer solution, and containers for buffer solution. etc.
Operation Steps
1. Put the gel with glass plates into the transfer buffer solution
2. Measure the gel size
3. Prepare 3 pieces of filter paper according the gel size, and the size of the filter paper should be a little bigger than the gel size; Here we use Whatman filter paper;
4. Put 3 pieces of filter paper into buffer solution slowly, and let the filter paper immerse into the buffer totally, and avoid to have air bubbles;
5. Prepare and cut nitrocellulose membrane according to the size of the gel and filter paper; the size of nitrocellulose membrane should be bigger than the size of the gel and filter paper;
6. Put the nitrocellulose membrane into the buffer solution;
7. Take out the 3 pieces filter paper and drop away extra buffer solution until no buffer solution drop from the membrane; and then place the filter paper on the bottom of DYCP-40C;
8. Take the gel from glass plates, gently clean the stacking gel, and put the gel into the buffer solution;
9. Place the gel on the filter paper, start from one end of the gel to avoid the air bubbles;
10. Use a proper tool to take away the air bubbles between the gel and filter paper.
11. Cover the nitrocellulose membrane on the gel, the rough side toward the gel. And then use a proper tool to take away the air bubbles between the membrane and the gel. Put 3 pieces of filter paper on the membrane. Still need to use a proper tool to take away the air bubbles between the filter paper and the membrane.
12. Cover the lid, and set the electrophoresis running parameters, the constant current 80mA;
13. The electrophoresis is done. We get the result as following;
Beijing Liuyi Biotechnology Co., Ltd has been specialized in manufacturing electrophoresis products for more than 50 years. We are ISO9001 & ISO13485 certified company and specializing in manufacturing electrophoresis tanks, power supplies, UV transilluminator, and gel documentation & analysis system, meanwhile, we provide OEM service for our customers, as well as ODM service.
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Post time: Apr-04-2023