Escherichia coli expressed exogenous proteins, which were suspended in PBS containing 1% triton-x-100 after ultrasonic treatment. The ultrasonic effect was better, and the effect of 1% triton-x-100 was still obvious. Some bacteria are also effective.
Escherichia coli expressed exogenous proteins, which were suspended in PBS containing 1% triton-x-100 after ultrasonic treatment. The ultrasonic effect was better, and the effect of 1% triton-x-100 was still obvious. Some bacteria are also effective.
Bacterial precipitation was directly added to sample 1b buffer, 5ul mercaptoethanol was added, mixed, centrifugated, boiled for 10min, directly loaded, dyed and decolorized in the following steps: put the glue into the appropriate amount of microwave dyeing solution for 1min, and replace a large amount of dyeing solution. The water can be boiled in the microwave for ten minutes.
After the expression of the recombinant protein, the cells were broken with ultrasonic wave, and 400w ice bath was used to break 2s and 1s, but a large amount of foam was generated within a short time, affecting the destructive power. Ultrasonic cell breaker. Both PBS and tris buffers do this. It's not completely broken, and the target protein is present in these unbroken cells.
1. Bubbles will be generated because the probe position is not set. The probe must be close to the bottom, about 25px. The power varies depending on the damage to the ultrasonic battery, but you can look at the liquid level, which fluctuates but not too strongly.
2, break 3S, stop 10S, break 20 or 30 times.
3. The position of the horn should also be paid attention to. If the sound is wrong, it should be adjusted in time. In addition, from the point of view of the concentration of bacteria can be considered. Increase the volume as much as possible, the strength should not exceed 60%.
4. Try to stop 8s for 8s. For some bacterial proteins, it is difficult for your method to dissipate heat, leading to protein denaturation and bubbles.
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