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Ac-DEVD-pNA *CAS 189950-66-1*
Ac-DEVD-pNA is a colorimetric substrate for caspase 3, a protease that is rapidly activated when cells are exposed to apoptotic conditions and that cleaves poly(ADP-ribose) polymerase.
Example protocol
AT A GLANCE
Important notes
It is important to store at <-15 °C and should be stored in cool, dark place.
It can be used within 12 months from the date of receipt.
PREPARATION OF STOCK SOLUTION
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
Ac-DEVD-pNA stock solution (100 mM):
Add 80 µL of DMSO to 5 mg to make Ac-DEVD-pNA stock solution. Note: The stock solution should be used promptly; any unused solution should be aliquoted and refrozen at -20 o C. Avoid repeated freeze-thaw cycles and keep from light.
PREPARATION OF WORKING SOLUTION
Ac-DEVD-pNA working solution (0.2 mM):
Add assay buffer of your choice to Ac-DEVD-pNA stock solution to make working solution. Note: Buffer can be used such as 50 mM Tris-HCl, pH 7.5, 0.3% NP-40, 1.0 mM DTT)
SAMPLE EXPERIMENTAL PROTOCOL
- Add 0.01 mL of enzyme solution (or cell lysate*) into 0.09 mL assay solution (from Step 2) to make a final volume of 0.1 mL. Note: Lyse cells in 50 mM Tris-HCl, pH 7.5, 0.3% NP-40, 1.0 mM DTT, at a density of 2 x 106/mL).
- Incubate the assay solution at 37 °C for 0-3 hr. Take periodic readings of absorbance at 405 nm.
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of Ac-DEVD-pNA *CAS 189950-66-1* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
| 0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
| 1 mM | 156.597 µL | 782.987 µL | 1.566 mL | 7.83 mL | 15.66 mL |
| 5 mM | 31.319 µL | 156.597 µL | 313.195 µL | 1.566 mL | 3.132 mL |
| 10 mM | 15.66 µL | 78.299 µL | 156.597 µL | 782.987 µL | 1.566 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
| Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
| / | = | x | = |
Product family
| Name | Excitation (nm) | Emission (nm) |
| Ac-DEVD-AFC *CAS 201608-14-2* | 376 | 482 |
| Ac-DEVD-AMC *CAS 169332-61-0* | 341 | 441 |
| Ac-DEVD-aminoluciferin | 362 | 499 |
Citations
View all 1 citations: Citation Explorer
Human high temperature requirement serine protease A1 (HTRA1) degrades tau protein aggregates
Authors: Tennstaedt, Annette and Pöpsel, Simon and Truebestein, Linda and Hauske, Patrick and Brockmann, Anke and Schmidt, Nina and Irle, Inga and Sacca, Barbara and Niemeyer, Christof M and Br, undefined and t, Rol and , undefined and others, undefined
Journal: Journal of Biological Chemistry (2012): 20931--20941
Authors: Tennstaedt, Annette and Pöpsel, Simon and Truebestein, Linda and Hauske, Patrick and Brockmann, Anke and Schmidt, Nina and Irle, Inga and Sacca, Barbara and Niemeyer, Christof M and Br, undefined and t, Rol and , undefined and others, undefined
Journal: Journal of Biological Chemistry (2012): 20931--20941
Page updated on November 20, 2024
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