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Index / Biovis / Caspase-5 Substrate WEHD-pNA; Appearance Liquid / Product Detail : 1102-200 Caspase-5 Substrate WEHD-pNA; Appearance Liquid
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Aug
30th

#1102-200 Caspase-5 Substrate WEHD-pNA; Appearance Liquid

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  Price : 379   EUR
431   USD
294   GBP
1595   Zloty
50814   JPY
2928   NOK
3138   SEK
429   CHF

Product name : Caspase-5 Substrate WEHD-pNA; Appearance Liquid

Catalog number : 1102-200

Quantity: 200 assays

Availability: Yes

Supplier name : Biovis

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More Details about

STORAGE: Store at –20°C, protected from light.
SHELF LIFE: 1 year under proper storage conditions
MOL. WEIGHT: 747.7
SEQUENCE: Ac-Trp-Glu-His-Asp-pNA
PURITY: >99% by HPLC analysis.


DESCRIPTION:
Ready-to-use colorimetric substrate for caspase-5 and related caspases that recognize the amino acid sequence WEHD. Caspase-5 and related caspase activity can be quantified by spectrophotometric detection of free pNA ( = 400 nm) after cleaved from the peptide substrate WEHD-pNA, using a spectrophotometer or multi-well plate reader. The ready-to-use caspase substrate provides an economic alternative for researchers who perform large volume of caspase assays.


ASSAY PROCEDURE:
1. Induce apoptosis in cells by desired method. Concurrently incubate a control culture without induction.
2. Count cells and pellet 1-5 x 106 cells.
3. Resuspend cells in 50 μl of chilled Cell Lysis Buffer (Cat.# 1067-100) and incubate cells on ice for 10 minutes.
4. Centrifuge for 1 min in a microcentrifuge (10,000 x g).
5. Transfer supernatant (cytosolic extract) to a fresh tube and put on ice.
6. Assay protein concentration.
7. Dilute 50-200 μg protein to 50 μl Cell Lysis Buffer for each assay.
8. Add 50 μl of 2X Reaction Buffer (Cat.# 1068-20, -80) containing 10 mM DTT (Cat.# 1201-1) to each sample.
9. Add 5 μl of the 4 mM of WEHD-pNA (200 μM final conc.) and incubate at 37°C for 1-2 hour.
10. 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 (Cat.#1066-100, -500) and using regular cuvette (note: Dilution of the samples proportionally decreases the reading).
You may also perform the entire assay directly in a 96-well plate.
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.


For Research Use Only! Not to be used in humans.

 

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