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Nanyang Technological University

Protein Extraction from Neuronal Cells

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Transkript

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.

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Protein Extraction from Neuronal Cells

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