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ALX-850-220 Revised 22-Apr-08
Caspase-6 Fluorometric Assay Kit
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PRODUCT LINE Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY Caspase Kits & Sets
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-850-220-KI01   25 tests 175.00 USD Add To Cart
Product Specification
KIT/SET CONTAINS: 25ml Cell Lysis Buffer, 2ml 2X Reaction Buffer, 125µl VEID-AFC Substrate (1mM), 100µl DTT (1M).
SHIPPING: SHIPPED ON BLUE ICE
LONG TERM STORAGE: -20°C
HANDLING: Protect from light. Store Cell Lysis Buffer and 2X Reaction Buffer at 4°C after opening.
Product Description
Based on the detection of AFC (7-amino-4-trifluoromethyl coumarin) after cleavage from the labelled substrate VEID-AFC.
General Information
VEID-AFC emits blue light (λmax = 400nm); upon cleavage of the substrate by caspase-6 or related caspases, free AFC emits a yellow-green fluorescence (λmax = 505nm), which can be quantified using a fluorometer or a fluorecence microtiter plate reader. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-6 activity.
MANUFACTURER

Manufactured by BioVision.
Further Categories Containing This Product:
Activity Assays
 
 
ALX-201-040 Revised 24-Jan-05
Caspase-7 (active) (human) (recombinant)
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PRODUCT LINE Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY Caspases
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Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-040-C005   5 µg 255.00 USD Add To Cart
Product Specification
EC: 3.4.22.-
SOURCE/HOST: Produced in E. coli.
FORMULATION: Liquid. 5µg in 25µl of 50mM TRIS (pH 8.0) containing 100mM sodium chloride and 50mM imidazole.
SHIPPING: SHIPPED ON DRY ICE
LONG TERM STORAGE: -80°C
USE/STABILITY: Stable at least one week when stored at +4°C.
HANDLING: Avoid freeze/thaw cycles.
Product Specific Literature References
[1] Biochemical characteristics of caspases-3, -6, -7, and -8: H.R. Stennicke & G.S. Salvesen; J. Biol. Chem. 272, 25719 (1997) Abstract; Full Text
General Information
When expressed in E. coli, caspase-7 spontaneously undergoes autoprocessing to yield the subunits characteristic of the active enzyme.
The rate of caspase-7 enzymatic hydrolysis can be measured by the release of AMC from the caspase substrate Ac-DEVD-AMC (Prod. No. ALX-260-031) as emission at 440nm and excitation at 380nm using a spectrofluorometer [1].
BACKGROUND/TECHNICAL INFORMATION
Enzyme Evaluation Protocol
This protocol is used to measure caspase enzyme activity [1]. The synthetic fluorogenic peptide Ac-DEVD-AMC (Prod. No. ALX-260-031) is used as the caspase enzyme substrate. The enzyme cleaves the substrate releasing the fluorescent AMC (7-amino-4-methylcoumarin). AMC release can be measured by spectrofluorometry using UV.

1. Add 20mM of Ac-DEVD-AMC (Prod. No. ALX-260-031) to 1ml of assay buffer (20mM PIPES, 100mM sodium chloride, 10mM DTT, 1mM EDTA, 0.1% (w/v) CHAPS, 10% sucrose, pH 7.2). Add the appropriate amount of  active caspase to the mixture (50ng/ml).
2. Incubate for 1 hour at 37°C.
3. Measure the AMC liberated from the Ac-DEVD-AMC using a spectrofluorometer with an excitation wavelength of 380nm and an emission wavelength of 440nm.

Note: Enzyme activity can also be measured using the caspase enzyme substrate Ac-DEVD-pNA (Prod. No. ALX-260-033) at 37°C in a spectromax at 405nm wavelength.

Swiss-Prot link P55210: Caspase-7 (human) (precursor)
AfCS Signalling Gateway link A000502: Caspase-7 (mouse)
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-061 Revised 21-Jul-08
Caspase-7 (active) (human) (recombinant)
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PRODUCT LINE Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY Caspases
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-061-U025   25 U 203.00 USD Add To Cart
ALX-201-061-U100   100 U 371.00 USD Add To Cart
Product Specification
EC: 3.4.22.60
SOURCE/HOST: Produced in E. coli.
PURITY: ≥90% (SDS-PAGE)
FORMULATION: Lyophilized.
RECONSTITUTION: Reconstitute to 1U/µl with PBS containing 15% glycerol.
SPECIFIC ACTIVITY: ~25’000U/mg protein. One unit is defined as the amount of enzyme that cleaves 1nmol of the caspase substrate DEVD-pNA (pNA: p-nitroanaline) per hour at 37°C in a reaction solution containing 50mM HEPES, pH 7.2, 50mM NaCl, 0.1% CHAPS, 10mM EDTA, 5% glycerol and 10mM DTT.
APPLICATION: Useful in screening caspase inhibitors, studying enzyme regulation, determining specificity of caspase substrates or as positive control in caspase activity assays. We recommend using 1 unit per assay for analyzing caspase activity.
SHIPPING: SHIPPED ON DRY ICE
LONG TERM STORAGE: -80°C
HANDLING: Avoid freeze/thaw cycles. After reconstitution, prepare aliquots and store at -80°C.
General Information
BACKGROUND/TECHNICAL INFORMATION Swiss-Prot link P55210: Caspase-7 (human) (precursor)
AfCS Signalling Gateway link A000502: Caspase-7 (mouse)
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-041 Revised 03-Feb-05
Caspase-8 (active) (human) (recombinant)
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PRODUCT LINE Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY Caspases
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-041-C005   5 µg 255.00 USD Add To Cart
Product Specification
EC: 3.4.22.-
SOURCE/HOST: Produced in E. coli.
FORMULATION: Liquid. 5µg in 25µl of 50mM TRIS (pH 8.0) containing 100mM sodium chloride and 50mM imidazole.
SHIPPING: SHIPPED ON DRY ICE
LONG TERM STORAGE: -80°C
USE/STABILITY: Stable for at least one week when stored at +4°C.
HANDLING: Avoid freeze/thaw cycles.
Product Specific Literature References
Biochemical characteristics of caspases-3, -6, -7 and -8: H.R. Stennicke & G.S. Salvesen; J. Biol. Chem. 272, 25719 (1997) Abstract; Full Text
General Information
When expressed in E. coli, caspase-8 spontaneously undergoes autoprocessing to yield the subunits characteristic of the active enzyme.
The rate of caspase-8 enzymatic hydrolysis can be measured by the release of AMC from the caspase substrate Ac-DEVD-AMC (Prod. No. ALX-260-031) as emission at 440nm and excitation at 380nm using a spectrofluorometer.
BACKGROUND/TECHNICAL INFORMATION
Enzyme Evaluation Protocol
This protocol is used to measure caspase enzyme activity [1]. The synthetic fluorogenic peptide Ac-DEVD-AMC (Prod. No. ALX-260-031) is used as the caspase enzyme substrate. The enzyme cleaves the substrate releasing the fluorescent AMC (7-amino-4-methylcoumarin). AMC release can be measured by spectrofluorometry using UV.

1. Add 20mM of Ac-DEVD-AMC (Prod. No. ALX-260-031) to 1ml of assay buffer (20mM PIPES, 100mM sodium chloride, 10mM DTT, 1mM EDTA, 0.1% (w/v) CHAPS, 10% sucrose, pH 7.2). Add the appropriate amount of  active caspase to the mixture (50ng/ml).
2. Incubate for 1 hour at 37°C.
3. Measure the AMC liberated from the Ac-DEVD-AMC using a spectrofluorometer with an excitation wavelength of 380nm and an emission wavelength of 440nm.

Note: Enzyme activity can also be measured using the caspase enzyme substrate Ac-DEVD-pNA (Prod. No. ALX-260-033) at 37°C in a spectromax at 405nm wavelength.

Swiss-Prot link Q14790: Caspase-8 (human) (precursor)
AfCS Signalling Gateway link A000503: Caspase-8 (mouse)
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-062 Revised 04-Aug-08
Caspase-8 (active) (human) (recombinant)
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PRODUCT LINE Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY Caspases
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-062-U025   25 U 203.00 USD Add To Cart
ALX-201-062-U100   100 U 371.00 USD Add To Cart
Product Specification
EC: 3.4.22.-
SOURCE/HOST: Produced in E. coli.
PURITY: ≥95% (SDS-PAGE)
FORMULATION: Lyophilized.
RECONSTITUTION: Reconstitute to 1U/µl with PBS containing 15% glycerol.
SPECIFIC ACTIVITY: 5’000U/mg protein. One unit is defined as the amount of enzyme that cleaves 1nmol of the caspase substrate IETD-pNA per hour at 37°C in a reaction solution containing 50mM HEPES, pH 7.2, 50mM NaCl, 0.1% CHAPS, 10mM EDTA, 5% glycerol and 10mM DTT.
APPLICATION: Useful in screening caspase inhibitors, studying enzyme regulation, determining specificity of caspase substrates or as positive control in caspase activity assays. We recommend using 1U per assay for analyzing caspase activity.
SHIPPING: SHIPPED ON DRY ICE
LONG TERM STORAGE: -80°C
HANDLING: Avoid freeze/thaw cycles. After reconstitution, prepare aliquots and store at -80°C.
Product Images
Please click on thumbnails to enlarge.
General Information
Caspase-8 is a member of the interleukin-1β converting enzyme (ICE) family of cysteine proteases. Same as other caspases, caspase-7 also exists in cells as an inactive proenzyme. During apoptosis procaspase-8 is processed at aspartate residues by self-proteolysis and/or cleavage by another caspase. The processed form of caspase-8 consists of large and small subunits which associate to form the active enzyme.
BACKGROUND/TECHNICAL INFORMATION Swiss-Prot link Q14790: Caspase-8 (human) (precursor)
AfCS Signalling Gateway link A000503: Caspase-8 (mouse)
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-522-054 Revised 24-Mar-05
Caspase-8 (active) (human) (recombinant) (high purity)
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SYNONYMS FLICE (active) (human) (recombinant) (high purity)
Mach-1 (active) (human) (recombinant) (high purity)
Mch5 (active) (human) (recombinant) (high purity)
PRODUCT LINE Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY Caspases
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ALX-522-054-C010   10 µg 180.00 USD Add To Cart
Product Specification
MW: ~29kDa; α-subunit: ~18kDa; β-subunit: ~11kDa.
EC: 3.4.22.-
SOURCE/HOST: Produced in E.coli.
CONCENTRATION: 0.1mg/ml.
PURITY: ≥95% (SDS-PAGE)
FORMULATION: Liquid. In 100mM HEPES, pH 7.5, containing 4mM DTT, 10% sucrose, 0.1% CHAPS.
SPECIFIC ACTIVITY: ~2'000U/mg. One unit is defined as the amount of enzyme that cleaves 1nmol of the caspase substrate Ac-DEVD-AMC (Prod. No. ALX-260-031) per minute at 37°C in a buffered solution.
SHIPPING: SHIPPED ON DRY ICE
LONG TERM STORAGE: -80°C
USE/STABILITY: Stable for at least 1 year after receipt if stored at -80°C.
HANDLING: Avoid freeze/thaw cycles.
Product Images
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Product Specific Literature References
Insights into the regulatory mechanism for caspase-8 activation  : M. Donepudi, et al. ; Mol. Cell 11, 543 (2003) Abstract
General Information
BACKGROUND/TECHNICAL INFORMATION The rate of recombinant human caspase-8 enzymatic activity can be measured by the release of AMC from the Ac-DEVD-AMC caspase substrate as emission at 440nm upon excitation at 380nm using a spectrofluorometer (Stennicke and Salvesen 1997).

Swiss-Prot link Q14790: Caspase-8 (human) (precursor)
AfCS Signalling Gateway link A000503: Caspase-8 (mouse)
MANUFACTURER Manufactured by Apotech Corporation.
General Literature References
Purification and catalytic properties of human caspase family members: M. Garcia-Calvo, et al.; Cell Death Differ. 6, 362 (1999) Abstract
Biochemical characteristics of caspases-3, -6, -7, and -8: H.R. Stennicke & G.S. Salvesen; J. Biol. Chem. 272, 25719 (1997) Abstract
Caspases: preparation and characterization: H.R. Stennicke & G.S. Salvesen; Methods 17, 313 (1999) Abstract
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-163 Revised 04-Mar-05
Caspase-8 (active) (mouse) (recombinant)
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PRODUCT LINE Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY Caspases
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-163-C020   20 µg 280.00 USD Add To Cart
Product Specification
SOURCE/HOST: Produced in E. coli.
CONCENTRATION: 1mg/ml
PURITY: ~90% (SDS-PAGE)
FORMULATION: Liquid. In 50mM HEPES, pH 7.4, containing 100mM NaCl, 0.5% CHAPS, 10mM DTT, 1mM EDTA and 50% glycerol.
SPECIFIC ACTIVITY: ≥100U/µg protein. One unit is defined as the amount of enzyme that cleaves 1nmol of the caspase substrate Z-IETD-AMC (AMC: 7-aminomethylcoumarin) (Prod. No. ALX-260-042) per hour at +30°C in a reaction solution containing 20µM Z-IETD-AMC, 50mM HEPES, pH 7.4, 100mM NaCl, 0.5% CHAPS, 10mM DTT, 1mM EDTA and 10% glycerol.
SHIPPING: SHIPPED ON DRY ICE
SHORT TERM STORAGE: -20°C
LONG TERM STORAGE: -80°C
USE/STABILITY: Stability at -20°C: 1-2 weeks.
HANDLING: Avoid freeze/thaw cycles. After opening, prepare aliquots and store at -80°C.
General Information
BACKGROUND/TECHNICAL INFORMATION Note: Activity of this enzyme may essentially decrease after repeated freezing and thawing. For consistent results, it is recommended to use fresh aliquots.
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-850-221 Revised 22-Apr-08
Caspase-8 Colorimetric Assay Kit
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PRODUCT LINE Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY Caspase Kits & Sets
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-850-221-KI01   25 tests 175.00 USD Add To Cart
Product Specification
KIT/SET CONTAINS: 25ml Cell Lysis Buffer, 2ml 2X Reaction Buffer, 125µl IETD-pNA Substrate (4mM), 100µl DTT (1M), 25ml Dilution Buffer.
SHIPPING: SHIPPED ON BLUE ICE
LONG TERM STORAGE: -20°C
HANDLING: Protect from light. Store Cell Lysis Buffer, 2X Reaction Buffer and Dilution Buffer at 4°C after opening.
Product Description
Based on the spectrophotometric detection of the chromophore pNA (p-nitroaniline) after cleavage from the labelled substrate IETD-pNA.
General Information
The pNA light emission can be quantified using a spectrophotometer or a microtiter plate reader at 400 or 405nm. Comparison of the absorbance of pNA from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-8 activity.
MANUFACTURER Manufactured by BioVision.
Further Categories Containing This Product:
Activity Assays
 
 
ALX-850-222 Revised 21-Apr-08
Caspase-8 Fluorometric Assay Kit
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PRODUCT LINE Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY Caspase Kits & Sets
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-850-222-KI01   25 tests 175.00 USD Add To Cart
Product Specification
KIT/SET CONTAINS: 25ml Cell Lysis Buffer, 2ml 2X Reaction Buffer, 125µl IETD-AFC Substrate (1mM), 100µl DTT (1M).
SHIPPING: SHIPPED ON BLUE ICE
LONG TERM STORAGE: -20°C
HANDLING: Protect from light. Store Cell Lysis Buffer and 2X Reaction Buffer at 4°C after opening.
Product Description
Based on the detection of AFC (7-amino-4-trifluoromethyl coumarin) after cleavage from the labelled substrate IETD-AFC.
General Information
IETD-AFC emits blue light (λmax = 400nm); upon cleavage of the substrate by caspase-8 or related caspases, free AFC emits a yellow-green fluorescence (λmax = 505nm), which can be quantified using a fluorometer or a fluorecence microtiter plate reader. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-8 activity.
MANUFACTURER Manufactured by BioVision.
Further Categories Containing This Product:
Activity Assays
 
 
ALX-201-047 Revised 14-Jul-08
Caspase-9 (active) (human) (recombinant)
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PRODUCT LINE Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY Caspases
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-047-U025   25 U 231.00 USD Add To Cart
ALX-201-047-U100   100 U 399.00 USD Add To Cart
Product Specification
EC: 3.4.22.62
SOURCE/HOST: Produced in E. coli.
PURITY: ≥90% (SDS-PAGE)
FORMULATION: Lyophilized.
RECONSTITUTION: Reconstitute to 1U/µl with 25µl or 100µl PBS containing 15% glycerol.
Note: It is normal to see some insoluble protein following reconstitution. This is mainly due to denaturation of small amounts of protein during preparation. Precipitate should not affect activity. If preferred, remove precipitate by brief centrifugation.
SPECIFIC ACTIVITY: ~400U/mg protein. One unit is defined as the amount of enzyme that cleaves 1nmol of the caspase substrate LEHD-pNA per hour at 37°C in a reaction solution containing 50mM HEPES, pH 7.2, 50mM NaCl, 0.1% CHAPS, 10mM EDTA, 5% glycerol and 10mM DTT.
APPLICATION: Useful in screening caspase inhibitors, studying enzyme regulation, determining target substrates or as positive control in caspase activity assays. We recommend using 1 unit per assay for analyzing caspase activity.
SHIPPING: SHIPPED ON DRY ICE
LONG TERM STORAGE: -80°C
HANDLING: Avoid freeze/thaw cycles. After reconstitution, prepare aliquots and store at -80°C.
Product Specific Literature References
The Ced-3/interleukin 1beta converting enzyme-like homolog Mch6 and the lamin-cleaving enzyme Mch2alpha are substrates for the apoptotic mediator CPP32: S.M. Srinivasula, et al.; J. Biol. Chem. 271, 27099 (1996) Abstract; Full Text
Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade: P. Li, et al.; Cell 91, 479 (1997) Abstract
A conserved XIAP-interaction motif in caspase-9 and Smac/DIABLO regulates caspase activity and apoptosis: S.M. Srinivasula, et al.; Nature 410, 112 (2001) Abstract
ICE-LAP6, a novel member of the ICE/Ced-3 gene family, is activated by the cytotoxic T cell protease granzyme B: H. Duan, et al.; J. Biol. Chem. 271, 16720 (1996) Abstract; Full Text
Caspase-9, Bcl-XL, and Apaf-1 form a ternary complex: G. Pan, et al.; J. Biol. Chem. 273, 5841 (1998) Abstract; Full Text
General Information
Caspase-9 (also known as Ice-Lap6 and Mch6) is a member of the interleukin-1b converting enzyme (ICE) family of cysteine proteases. Similar as other caspases, caspase-9 also exists in cells as an inactive proenzyme. During the initiation of apoptosis procaspase-9 is processed at aspartate residues to form active caspase-9. As one of the initiator caspases, active caspase-9 functions to trigger the activation of downstream effector caspases, leading to disassembly of cell structures. The active recombinant caspase-9 is routinely tested for its ability to enzymatically cleave the caspase-9 colorimetric substrate Ac-LEHD-pNA (Prod. No. ALX-260-081).
BACKGROUND/TECHNICAL INFORMATION Swiss-Prot link P55211: Caspase-9 (human) (precursor)
AfCS Signalling Gateway link A000504: Caspase-9 (mouse)
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-136 Revised 14-Aug-08
Caspase-9 (active) (human) (recombinant)