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  • 產(chǎn)品名稱：CaspaseColorimetricSubstrateSet

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  • 產(chǎn)品廠商：Biovision
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CaspaseColorimetricSubstrateSet
詳情介紹：
Purpose Ready-to-use colorimetric substrates for members of caspase family proteases. All substrates were provided in liquid ready-to-use form.
Sample Type Cell Culture Cells
Detection Method Colorimetric
Characteristics Caspase-Family Colorimetric Substrate Set: Ready-to-Use Colorimetric Substrates for Assaying Activities of Caspase-1, -2, -3, -5, -6, -8 & -9 family proteases. Concentration: 4 mM.
Ready-to-use colorimetric substrates for members of caspase family proteases. All substrates were provided in liquid ready-to-use form. KIT CONTENTS: 125 μL (4 mM) each of the following substrates dissolved in DMSO: Caspase-1 Substrate, Ac-YVAD-pNA Caspase-2 Substrate, Ac-VDVAD-pNA Caspase-3 Substrate, Ac-DEVD-pNA Caspase-5 Substrate, Ac-WEHD-pNA Caspase-6 Substrate, Ac-VEID-pNA Caspase-8 Substrate, Ac-IETD-pNA Caspase-9 Substrate, Ac-LEHD-pNA
Components Caspase-1 Substrate, Ac-YVAD-pNA
Caspase-2 Substrate, Ac-VDVAD-pNA
Caspase-3 Substrate, Ac-DEVD-pNA
Caspase-5 Substrate, Ac-WEHD-pNA
Caspase-6 Substrate, Ac-VEID-pNA
Caspase-8 Substrate, Ac-IETD-pNA
Caspase-9 Substrate, Ac-LEHD-pNA
Alternative Name Caspase
Background Ready-to-use colorimetric substrates for members of caspase family proteases. All substrates were provided in liquid ready-to-use form.
Application Notes Detects specific Caspases involved in apoptosis within cells
Comment

Further details regarding sample type: Cell culture
Protocol 1. Induce apoptosis in cells by desired method. Concurrently incubate a control culture without induction.
2. Count cells and pellet 1-5 x 10^6 cells.
3. Resuspend cells in 50 μL of chilled Cell Lysis Buffer and incubate cells on ice for 10 minutes.
4. Centrifuge for 1 min in a microcentrifuge (10,000 x g).
5. Transfer supernatant to a fresh tube and assay protein Concentration.
6. Dilute 50-200 μg protein to 50 μL Cell Lysis Buffer for each assay.
7. Add 50 μL of 2X Reaction Buffer containing 10 mM DTT to each sample.
8. Add 5 μL of the 4 mM pNA conjugated substrates (200 μ M final conc.) into each tube individually and incubate at 37 °C for 1-2 hour.
9. Read samples at 400- or 405-nm in a microtiter plate reader, or spectrophotometer using a 100- μL micro quartz cuvet (Sigma), or dilute sample to 1 mL with Dilution Buffer and using regular cuvet (note: Dilution of the samples proportionally decreases the reading). Fold-increase in caspase activity can be determined by comparing these results with the level of the uninduced control. Note: Background reading from cell lysates and buffers should be subtracted from the readings of both induced and the uninduced samples before calculating fold increase in caspase activity.
Restrictions For Research Use only
Concentration 4 mM
Storage -20 °C
Expiry Date 6-12 months
Product cited in: Doostzadeh-Cizeron, Yin, Goodrich: "Apoptosis induced by the nuclear death domain protein p84N5 is associated with caspase-6 and NF-kappa B activation." in: The Journal of biological chemistry, Vol. 275, Issue 33, pp. 25336-41, 2000 (PubMed).