cOmplete™ Lysis-M Protocol & Troubleshooting

Product No. 04719956001

Protocol

Interference with immunoprecipitation

The sample does not have to be purified to remove this mild detergent. It does not interfere with immunoprecipitation, but is very effective in lysing mammalian cells.

Troubleshooting

cOmplete™ Lysis-M Inhibitor: Low protein concentration in lysate

Low protein concentration in cell lysates can indicate that the volume of lysis buffer is not sufficient to lyse cells. for this reason, it may not be possible to decrease the volume of the cOmplete inhibitor solution. For best results, use volumes recommended in the package insert. For a 6-well plate, use 200 to 400 μl per well. If the protein concentration in the lysate is still low, increase the incubation time and agitate the culture plate vigorously.

Be aware that low cell number could be the reason for low protein content. In this instance, it can be advantageous to scrape the cells from the plate and collect them by centrifugation. Remove as much of the supernatant as possible, and lyse the cell pellet in 50, 100, or 150 μl lysis buffer.

Materials

     

COMPLETE is a trademark of Roche

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