广州健仑生物科技有限公司

广州健仑生物科技有限公司

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李斯特菌检测卡（食品安全检测）

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8M dissolved in high concentrations of urea after centrifugation obtained inclusion body lysate, placed in a refrigerator after 4 degrees overnight precipitation, this phenomenon I have encountered in the experiment; began to feel very strange, and later found to be our refrigerator temperature control The problem was that the inter-laboratory temperature was at least 4 degrees below the set temperature but then I went down to SDS-PAGE and found that there was no adverse effect. So, do not worry, maybe your protein is just a joke on you!

Third, on the dissolution of inclusion bodies:

1, ask GST inclusion body dissolved

Directly with 8M urea or 6M guanidine melt, take the supernatant, measured concentration. After the direct dilution of the package is detected the corresponding antibody. (Since the rapid dilution corresponds to the refolding process, the product is used to detect no problems)

I used the inclusion body extraction method:

1. After PBS or saline wash induced bacterial cells, Buffer A resuspended bacteria, ultrasonic broken to clear

2.4 degrees, 10000rpm centrifuge for 10 minutes, the supernatant was discarded

3. The pellet is washed 2-3 times with PBS or normal saline

4. To the precipitate add 19.7ml Buffer A and 0.3ml 20% SKL (sodium lauryl sarcosine) stock solution, vigorously agitated, slowly dissolved, allowed to stand at room temperature for 30 minutes to 2 hours

5.4 degrees Centrifuge at 10000rpm for 10 minutes, take the supernatant

6. Add 20% PEG4000 210ul to a final concentration of 0.2%, add 50mM oxidized glutathione 420ul to a final concentration of 1mM, add 100mM glutathione 420ul to a final concentration of 2mM and let stand for 30 minutes To 2 hours (also 4 degrees refolding overnight)