Sie haben vollen Zugriff auf diesen Inhalt durch
Nanyang Technological University-- views • 1:16 min
Take a cell culture containing neuronal cancer cells attached to a coverslip and maintain it under cold conditions.
Remove the culture medium and repeatedly wash the cells with cold phosphate buffer to eliminate the residual medium.
Add a lysis buffer containing a low concentration of non-ionic detergent and protease inhibitors.
The non-ionic detergent disrupts the cell membrane to initiate lysis.
Meanwhile, the protease inhibitors block the cellular proteolytic enzymes, protecting the proteins from enzymatic degradation.
Next, use a cell scraper to dislodge the partially lysed cells from the coverslip.
Transfer the contents to a tube containing the lysis buffer.
Under cold conditions, use a homogenizer to mechanically lyse the cells, ensuring complete cell lysis and protein release.
Centrifuge the cell lysate.
Collect the supernatant into a fresh tube.
The extracted proteins from the neuronal tumor cells are now ready for further analysis.
Copyright © 2025 MyJoVE Corporation. Alle Rechte vorbehalten