Application of Ding Haoyuan Automatic Transfer Instrument Blotstainer in Western Blotting

Because the steps of protein immunoblotting are cumbersome and require high precision, the operation error in one step in the experiment will lead to waste of the whole experiment. And the primary anti-reagent is also more valuable, so the use of instrumentation can reduce the risk of the experiment and ensure the smooth progress of the experiment.

Dingyiyuan's fully automatic transfer device is a good alternative to manual operation. It is equipped with automatic pipetting system, temperature control system and incubator. The automatic pipetting system consists of 10 lines that are independent of each other to avoid cross-infection. The temperature control system ranges from 4 ° C to 80 ° C and satisfies the incubation temperature of the immunoblot and nucleic acid hybridization experiments. The entire system is automatically completed according to customer-defined operating procedures. In order to facilitate the use of the customer, the automatic transfer device is pre-set with an automatic procedure for immunoblotting, nucleic acid hybridization, in situ hybridization and immunohistochemistry.

The built-in standard protein immunoblotting procedure is as follows:

1, set the temperature to room temperature

2. Wash the membrane with 0.01 M PBS for 5 min × 2 times.

3, set the temperature to 10 ° C

4. Add the coating solution and shake gently for 1 hr.

5. Add primary antibody (diluted with 0.01 M PBS in the appropriate dilution ratio, the liquid must cover all of the membrane), 2 hr.

6. Discard the primary antibody and wash the membrane with 0.01 M PBS for 5 min × 4 times.

7. Add secondary antibody (horseradish peroxidase coupling) (diluted with 0.01 M PBS in appropriate dilution) for 0.5 hr.

8, set the temperature to room temperature

9. Discard the secondary antibody and wash the membrane with 0.01M PBS for 5 min × 10 times.
10. Add the coloring solution, avoid the light and develop color until the band appears. Put in the double distilled water to stop the reaction.

In the above-mentioned program, the automatic transfer device can realize the temperature control during the incubation of the protein immunoblotting , and further ensure the activity of the primary antibody, which is not achieved by the manual operation.

The entire program is automatically completed within 5 hours, eliminating the need for manual fluid changes and film removal. Of course, the above program can adjust the program of the automatic transfer device according to the actual needs of the user.