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The temperature is too high during electrophoresis

It is recommended to combine at 4 ° C overnight. 10) The antibody concentration is too high or the washing is not enough. It is recommended to reduce the antibody concentration and increase the number and time of washing. 11) The chemical coloring substrate is too much. It is recommended to add an appropriate amount of coloring base according to the instructions. Object 3. Why the shape of the band is not good? 1) The gelation is not uniform or the polymerization is not good. It is recommended to mix the solution thoroughly before pouring. 2) Some samples have a high salt concentration. It is recommended to remove the salt or adjust the sample salt concentration to be the same. 3) The buffer solution is old and the composition is changed. It can be reconstituted. 4)

There are air bubbles under the gel. Remove the air bubbles before electrophoresis. 5) The temperature is too high during electrophoresis, which can reduce the current or voltage. 6) The sample contains insoluble particles. It is recommended to stir the sample thoroughly. 4. Protein Is the band position (size) misaligned? The gel concentration is not correct. The position of the protein bands run by different concentrations of gel may be different.

You can adjust the concentration to inadequate antibody incubation. It is recommended to increase the antibody concentration and prolong the incubation time. 3) Enzyme inactivation, it is recommended to directly enzyme and substrate Mix and if the color does not develop, the enzyme is inactive. Select active enzyme conjugates within the validity period. 4) The target protein has post-translational modifications or splices. It is recommended to consult relevant literature to determine. 5) The sample does not contain the target protein or the target protein content is too low.

It is recommended to set a positive control comparison. As a result, the amount of sample is increased by 5. Is there a lot of banding? 1) The target protein has multiple modification sites, which can present multiple bands. It is recommended to consult the literature or perform bioinformatics analysis to obtain the modification site information of the protein sequence, and determine the actual size of the protein by demodification. 2) During sample processing Protease inhibitors are recommended for the degradation of the target protein.

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