We answered the question from the customer.
Here are some points that you need to pay particular attention to.

For user’s troubleshooting Ver2.33

Summary

  • For the passage of liquid factor through the filter, fill the passage (filter surface) with enough solution.
  • The recommended way to preprocess filters is washing with pure water and PBS after 1 minute with 100% ethanol and setting the filter to ICCP

Question:In the experiment with the filter, there is little passage of liquid factor?

Answer: Factors such as proteins and exosomes attach to filters, containers and extraction kits. Therefore, the more processes there are, the smaller the amount of recovery. If you can collect enough positive control samples (if there are no problems with the extraction process), first check the culture conditions and filter treatment. In most cases, the culture may be low or the filter may not be fully degassed. If the culture volume is small, the area of the filter contacting the culture solution will be small, and the co-culture effect may be significantly reduced.

In addition, air remains in the pores of the filter, so sufficient degassing is required before use. Please refer to the following pages for the specific amount, status, and filtering.