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Ac-DEVD-pNA *CAS 189950-66-1*

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Physical properties
Molecular weight638.58
SolventDMSO
Storage, safety and handling
Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12352200

OverviewpdfSDSpdfProtocol


See also: Caspases
CAS
189950-66-1
Molecular weight
638.58
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

  1. 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).

  2. 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 mg0.5 mg1 mg5 mg10 mg
1 mM156.597 µL782.987 µL1.566 mL7.83 mL15.66 mL
5 mM31.319 µL156.597 µL313.195 µL1.566 mL3.132 mL
10 mM15.66 µL78.299 µL156.597 µL782.987 µL1.566 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Citations


View all 1 citations: Citation Explorer
Human high temperature requirement serine protease A1 (HTRA1) degrades tau protein aggregates
Authors: Tennstaedt, Annette and P&ouml;psel, 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