User:Robwarden/Notebook/Caspase-3 Activatable Probe/2009/10/05

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Construct Luminescence Measurement

Induction of Apoptosis

Apoptosis was induced by 10μM staurosporine in half of the samples. Samples will be measured after 4hr and 6hr incubations at 37C. A Promega Luciferase Assay will be used.

4hr

Construct + -
1X Gaussia 4,143,449 19,643,584
5X Gaussia 1,283,243 8,496,529
1X Renilla 203,756,023 231
5X Renilla 220,700,532 1,302,498,176
No DNA 232 242
<bars title="Log10 Luminescence"

colors=336699,669933,993366 ylabel=10 xlabel legend grid=xy ymin=0 ymax=10 size=450x300> ,Apoptotic,Non-Apoptotic 1xG, 6.617361998, 7.293220728 5xG, 6.108308904, 6.929241544 1xR, 8.309110455, 2.36361198 5xR, 8.34380338, 9.114777124 No DNA, 2.365487985, 2.383815366 </bars>

6hr

Construct + -
1X Gaussia 397 26,226,718
5X Gaussia 1,755,862 9,029,151
1X Renilla 115,874,928 1,035,950,080
5X Renilla 112,831,944 568,635,136
No DNA 367 448
<bars title="Log10 Luminescence"

colors=336699,669933,993366 ylabel=10 xlabel legend grid=xy ymin=0 ymax=10 size=450x300> ,Apoptotic,Non-Apoptotic 1xG, 2.598790507, 7.418743947 5xG, 6.24449038, 6.955646916 1xR, 8.063989477, 9.015338828 5xR, 8.052432071, 8.754833691 No DNA, 2.564666064, 2.651278014 </bars>

Conclusions

  • Seems backwards
  • Perhaps enzymes are being degraded
  • Significant cell death was seen through microscope in both induced and uninduced samples