cOmplete™ Lysis-M EDTA-free Troubleshooting

Product No. 04719964001

Troubleshooting

Protein concentration in lysate

The low protein concentration in cell lysates may be an indication that the volume of lysis buffer is not sufficient to completely lyse the cells. In this case, do not decrease the volume of Complete inhibitor. For best results, use the volumes indicated in the package insert. For a 6-well plate, use 200-400 μl per well. If the protein concentration in the lysate is still low, increase the incubation time and shake the mixture more vigorously.

When there are low cell numbers in your primary cultures (because confluence is not reached), this can also be the reason for low protein content. to increase protein yield, scrape the cells from the plate and collect as many cells as possible using centrifugation. Remove as much of the supernatant as possible, and then find the optimum volume of Complete inhibitor by lysing the cell pellet in 50, 100, or 150 μl volumes.

Materials

     

COMPLETE is a trademark of Roche

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