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Index / Biovis / Caspase-12 Substrate ATAD-AFC; Appearance Liquid /Product Detail : 1117-1000 Caspase-12 Substrate ATAD-AFC; Appearance Liquid
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#1117-1000

Caspase-12 Substrate ATAD-AFC; Appearance Liquid

 Price: 1267   EUR
1439   USD
983   GBP

Product name : Caspase-12 Substrate ATAD-AFC; Appearance Liquid

Catalog number : 1117-1000

Quantity: 1000 assays

Availability: Yes

Supplier name : Biovis

Data sheet Data sheet : Ask more or other datasheet now !

More Details about

STORAGE: Store at –20°C, protected from light. Stable for 6 months
MOL. WEIGHT: 629.54
SEQUENCE: Ac-Ala-Thr-Ala-Asp-AFC (AFC, 7-amino-4-trifluoromethyl coumarin)
PURITY: >90% by HPLC analysis


DESCRIPTION:
Ready-to-use fluorometric substrate for caspases that recognize the amino acid sequence ATAD. Caspase activity can be quantified by fluorescent detection of free AFC after cleaved from the peptide substrate ATAD-AFC at Ex/Em = 400/505 nm using a fluorometer or a fluorescence plate reader. Alternatively, a shift in fluorescence from blue to green upon cleavage can be visualized using a hand-held long-UV lamp. The ready-to-use caspase substrate provides an economic alternative for researchers who perform large amount of caspase assays. Cell Lysis Buffer (Cat.# 1067-100), 2X Reaction Buffer (Cat.# 1068-20), DTT (Cat.# 1201-1), and other reagents used for caspase activity assays are also available separately.


ASSAY PROTOCOL:
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 or use 50-200 μg cell lysates if protein concentration has been measured.
3. Resuspend cells in 50 μl of chilled Cell Lysis Buffer (Cat.# 1067-100).
4. Incubate cells on ice for 10 minutes.
5. Add 50 μl of 2X Reaction Buffer (Cat.# 1068-20, -80) containing 10 mM DTT (Cat.# 1201-1) to each sample.
6. Add 5 μl of the 1 mM ATAD-AFC (50 μM final conc.) into each tube individually and incubate at 37°C for 1-2 hour.
7. Read samples in a fluorometer equipped with a 400-nm excitation filter and 505-nm emission filter. For a plate-reading set-up, transfer the samples to a 96-well plate. You may perform the entire assay directly in a 96-well plate.
Fold-increase in ATAD-dependent caspase activity can be determined by comparing these results with the level of the uninduced control.

 

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