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Citações e referências (12)
Invitrogen™
EnzChek™ Caspase-3 Activity Assay Kits
Detect apoptosis in cell extracts and purified enzyme preparations with EnzChek Caspase-3 Activity Assay Kits with Z-DEVD-AMC substrate or Z-DEVD-R110 substrates.
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| Número do catálogo | Label or Dye |
|---|---|
| E13184 | Rhodamine 110 |
| E13183 | AMC (7-amino-4-methylcoumarin) |
Número do catálogo E13184
Preço (BRL)
4.365,38
Each
Label or Dye:
Rhodamine 110
Preço (BRL)
4.365,38
Each
Enable simple and reliable detection of caspase-3-activity-mediated apoptosis with the EnzChek Caspase-3 Activity Assay Kit, which takes advantage of specially designed substrates that fluoresce upon cleavage by the caspase-3 enzyme. The EnzChek Caspase-3 Activity Assay Kit is available with either the Z-DEVD-AMC or Z-DEVD-R110 substrate, which yields a bright fluorescent product upon proteolytic cleavage by caspase 3/7.
The EnzChek Caspase-3 Activity Assay Kit enables detection of apoptosis by providing a simple and reliable method for assaying caspase-3/7 activity. The kit can be used to continuously measure the activity of caspase-3/7 in cell extracts and purified enzyme preparations using a fluorometer or fluorescence microplate reader.
Key features of the EnzChek Caspase-3 Activity Assay Kit includes:
• Fluorescent assay using standard fluorescein (FITC) excitation/emission settings
• Format allows for continuous measurement of caspase-3/7 activity in cell extracts
• Fluorescent and enzymatic controls included
The aminomethylcoumarin (AMC)-derived substrate (Z-aspartic acid-glutamic acid-valine-aspartic acid (DEVD)-AMC) that is included in the EnzChek Caspase-3 Activity Assay Kit is weakly fluorescent in the UV range (excitation/emission maxima ˜330/390 nm), but yields a bright, blue-fluorescent product (excitation/emission maxima ˜342/441 nm) upon proteolytic cleavage. The kit can be used to continuously measure the activity of caspase-3 and closely related proteases in cell extracts and purified enzyme preparations using a fluorometer or fluorescence microplate reader.
The rhodamine 110-derived substrate (Z-DEVD-R110) used in the EnzChek Caspase-3 Activity Assay Kit is a non-fluorescent bisamide compound that, upon enzymatic cleavage, is converted in a two-step process to the fluorescent monoamide and then to the even more fluorescent R110 product. Both of these hydrolysis products exhibit spectral properties similar to those of fluorescein, with peak excitation and emission wavelengths of 496 nm and 520 nm, respectively.
In addition to the substrates, the EnzChek Caspase Activity Assay Kit contains the reversible aldehyde inhibitor Ac-DEVD-CHO, as well as an AMC or R110 reference standard. The Ac-DEVD-CHO inhibitor confirms that the fluorescence signals in both induced and control cell populations are due to the activity of the caspase-3/7 protease. The reference standard allows for quantification of the amount of AMC or R110 released in the reaction.
Key features of the EnzChek Caspase-3 Activity Assay Kit includes:
• Fluorescent assay using standard fluorescein (FITC) excitation/emission settings
• Format allows for continuous measurement of caspase-3/7 activity in cell extracts
• Fluorescent and enzymatic controls included
The aminomethylcoumarin (AMC)-derived substrate (Z-aspartic acid-glutamic acid-valine-aspartic acid (DEVD)-AMC) that is included in the EnzChek Caspase-3 Activity Assay Kit is weakly fluorescent in the UV range (excitation/emission maxima ˜330/390 nm), but yields a bright, blue-fluorescent product (excitation/emission maxima ˜342/441 nm) upon proteolytic cleavage. The kit can be used to continuously measure the activity of caspase-3 and closely related proteases in cell extracts and purified enzyme preparations using a fluorometer or fluorescence microplate reader.
The rhodamine 110-derived substrate (Z-DEVD-R110) used in the EnzChek Caspase-3 Activity Assay Kit is a non-fluorescent bisamide compound that, upon enzymatic cleavage, is converted in a two-step process to the fluorescent monoamide and then to the even more fluorescent R110 product. Both of these hydrolysis products exhibit spectral properties similar to those of fluorescein, with peak excitation and emission wavelengths of 496 nm and 520 nm, respectively.
In addition to the substrates, the EnzChek Caspase Activity Assay Kit contains the reversible aldehyde inhibitor Ac-DEVD-CHO, as well as an AMC or R110 reference standard. The Ac-DEVD-CHO inhibitor confirms that the fluorescence signals in both induced and control cell populations are due to the activity of the caspase-3/7 protease. The reference standard allows for quantification of the amount of AMC or R110 released in the reaction.
For Research Use Only. Not for use in diagnostic procedures.
Especificações
DescriptionEnzChek Caspase-3 Assay Kit #2, Z-DEVD-R110 substrate
Excitation/Emission496/520 (cleaved substrate)
For Use With (Equipment)Fluorometer, Microplate Reader
Label TypeOther Label(s) or Dye(s)
Label or DyeRhodamine 110
No. of Reactions500 Assays (reaction volume 100μL per assay)
Product LineEnzChek
Product TypeCaspase Assay Kit
Quantity500 Assays
Shipping ConditionRoom Temperature
Storage RequirementsStore in freezer (-5 to -30°C) and protect from light.
Detection MethodFluorescence
FormatCuvettes, 96-well Plate
Unit SizeEach
Frequently asked questions (FAQs)
Is the Z-DEVD-R110 substrate in the EnzChek Caspase-3 Assay Kit #2 cell permeable?
Citações e referências (12)
Citações e referências
Abstract
Novel small molecule inhibitors of caspase-3 block cellular and biochemical features of apoptosis.
Journal:J Pharmacol Exp Ther
PubMed ID:12490620
'Caspase-3 is an intracellular cysteine protease, activated as part of the apoptotic response to cell injury. Its interest as a therapeutic target has led many to pursue the development of inhibitors. To date, only one series of nonpeptidic inhibitors have been described, and these have limited selectivity within the caspase
Role of the sphingosine rheostat in the regulation of cnidarian-dinoflagellate symbioses.
Journal:Biol Bull
PubMed ID:22186914
'The symbiosis between host cnidarians, such as corals and anemones, and their dinoflagellate symbionts is regulated by largely undescribed mechanisms that stabilize the symbiosis during normal conditions but lead to symbiosis breakdown, or cnidarian bleaching, during stress. Previous transcriptomic studies identified the sphingosine rheostat as a putative symbiosis regulatory pathway.
Fluorescent molecular probes V: a sensitive caspase-3 substrate for fluorometric assays.
Journal:Bioorg Med Chem Lett
PubMed ID:10576694
(Z-Asp-Glu-Val-Asp)-Rhodamine 110 [(Z-DEVD)2-Rh 110] was prepared and characterized as a sensitive fluorogenic substrate for the determination of caspase-3 activity.
Photodynamic therapy and cell death pathways.
Journal:Methods Mol Biol
PubMed ID:20552338
Photodynamic therapy (PDT) is the term used to describe the irradiation of photosensitized cells or tissue with phototoxic consequences. This process can result in the rapid initiation of not only apoptosis, an irreversible death pathway, but also autophagy. The procedures described here are designed to characterize the correlation between the
Rhodamine 110-linked amino acids and peptides as substrates to measure caspase activity upon apoptosis induction in intact cells.
Journal:Biochemistry
PubMed ID:10529236
Caspases (cysteine aspartate-specific proteases) are a structurally related group of cysteine proteases that cleave peptide bonds following specific recognition sequences. They play a central role in activating apoptosis of vertebrate cells. To measure apoptosis induced by various stimuli and at an early apoptotic stage, caspases are an ideal target. This