ALEXIS Biochemicals: Monoclonal Antibody to Caspase-3 (31A1067)

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ALX-201-047

Revised 14-Jul-08

Caspase-9 (active) (human) (recombinant)
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
ALX-201-047-U100		100 U	399.00 USD

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)
PRODUCT LINE	Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY	Caspases

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-136-C005		5 µg	255.00 USD

Product Specification

EC:	3.4.22.-
SOURCE/HOST:	Produced in E. coli.
CONCENTRATION:	0.2mg/ml
FORMULATION:	Liquid. In 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

A combinatorial approach defines specificities of members of the caspase family and granzyme B. Functional relationships established for key mediators of apoptosis: N.A. Thornberry, et al.; J. Biol. Chem. 272, 17907 (1997) Abstract; Full Text

General Information

When expressed in E. coli, caspase-9 spontaneously undergoes autoprocessing to yield the subunits characteristic of the active enzyme. The rate of caspase-9 enzymatic activity can be measured by the release of AMC from the caspase substrate Ac-LEHD-AMC (Prod. No. ALX-260-080) as emission at 440nm and excitation at 380nm using a spectrofluorometer.
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-850-223

Revised 22-Apr-08

Caspase-9 Colorimetric Assay Kit
PRODUCT LINE	Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY	Caspase Kits & Sets

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-850-223-KI01		25 tests	203.00 USD

Product Specification

KIT/SET CONTAINS:	25ml Cell Lysis Buffer, 2ml 2X Reaction Buffer, 125µl LEHD-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 LEHD-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 a treated sample with an untreated control allows determination of the fold increase in caspase-9 activity.
MANUFACTURER	Manufactured by BioVision.

Further Categories Containing This Product:

Activity Assays

ALX-850-224

Revised 22-Apr-08

Caspase-9 Fluorometric Assay Kit
PRODUCT LINE	Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY	Caspase Kits & Sets

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-850-224-KI01		25 tests	189.00 USD

Product Specification

KIT/SET CONTAINS:	25ml Cell Lysis Buffer, 2ml 2X Reaction Buffer, 125µl LEHD-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 LEHD-AFC.

General Information

LEHD-AFC emits blue light (λmax = 400nm); upon cleavage of the substrate by caspase-9 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-9 activity.

MANUFACTURER

Manufactured by BioVision.

Further Categories Containing This Product:

Activity Assays

ALX-201-137

Revised 16-Feb-05

Caspase-10 (active) (human) (recombinant)
PRODUCT LINE	Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY	Caspases

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-137-C005		5 µg	255.00 USD

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

Purification and catalytic properties of human caspase family members: M. Garcia-Calvo, et al.; Cell Death Differ. 6, 362 (1999) Abstract

General Information

When expressed in E. coli, caspase-10 spontaneously undergoes autoprocessing to yield the subunits characteristic of the active enzyme.
The rate of caspase-10 enzymatic activity can be measured by the release of AMC from the caspase substrate Ac-IETD-AMC (Prod. No. ALX-260-042) 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. The synthetic fluorogenic peptide Ac-IETD-AMC (Prod. No. ALX-260-042) 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-IETD-AMC (Prod. No. ALX-260-042) 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-IETD-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-IETD-pNA (Prod. No. ALX-260-045) at 37°C in a spectromax at 405nm wavelength.

Swiss-Prot link Q92851: Caspase-10 (human) (precursor)
AfCS Signalling Gateway link A000493: Caspase-10 (human)

Further Categories Containing This Product:

Enzymes • Recombinant Proteins/Fusion Proteins

ALX-201-091

Revised 20-Oct-05

Caspase-10/a (active) (human) (recombinant)
PRODUCT LINE	Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY	Caspases

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-091-U025		25 U	203.00 USD
ALX-201-091-U100		100 U	371.00 USD

Product Specification

EC:	3.4.22.-
SOURCE/HOST:	Produced in E. coli.
PURITY:	≥90% (SDS-PAGE)
FORMULATION:	Lyophilized.
RECONSTITUTION:	Reconstitute to 1U/µl with PBS containing 15% glycerol.
SPECIFIC ACTIVITY:	~8'000U/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.
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

Fas-associated death domain protein interleukin-1beta-converting enzyme 2 (FLICE2), an ICE/Ced-3 homologue, is proximally involved in CD95- and p55-mediated death signaling: C. Vincenz & V.M. Dixit; J. Biol. Chem. 272, 6578 (1997) Abstract; Full Text
In vitro activation of CPP32 and Mch3 by Mch4, a novel human apoptotic cysteine protease containing two FADD-like domains: T. Fernandes-Alnemri, et al.; PNAS 93, 7464 (1996) Abstract

General Information

Caspase-10 (also known as Mch4 and Flice2) is a member of the interleukin-1b converting enzyme (ICE) family of cysteine proteases. Similar as other caspases, caspase-10 exists in cells as an inactive proenzyme. During apoptosis, procaspase-10 is processed at aspartate residues by self-proteolysis and/or cleavage by upstream caspases. The processed form of caspase-10 consists of large (18kDa) and small (11kDa) subunits which associate to form the active enzyme.

BACKGROUND/TECHNICAL INFORMATION

Swiss-Prot link Q92851: Caspase-10 (human) (precursor)
AfCS Signalling Gateway link A000493: Caspase-10 (mouse)

Further Categories Containing This Product:

Enzymes • Recombinant Proteins/Fusion Proteins

ALX-201-193

Revised 15-Dec-04

Caspase-10/a (active) (human) (recombinant) (high stability)
PRODUCT LINE	Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY	Caspases

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-193-U050		50 U	320.00 USD

Product Specification

MW:	~17kDa (large subunit); ~12kDa (small subunit).
EC:	3.4.22.-
SOURCE/HOST:	Produced in E. coli.
CONCENTRATION:	~1mg/ml
PURITY:	≥95% (SDS-PAGE)
FORMULATION:	Liquid. 20µl in 100mM HEPES, pH 7.0, containing 10% sucrose, 10mM DTT, 2mM EDTA and 0.1% CHAPS.
SPECIFIC ACTIVITY:	~2'500U/mg protein. One unit is defined as the amount of enzyme that cleaves 1 nmol of the caspase substrate IETD-pNA per min. at 37°C in a buffered solution.
SHIPPING:	SHIPPED ON DRY ICE
LONG TERM STORAGE:	-80°C
USE/STABILITY:	Shows excellent freeze-thaw stability (see Figure).

Product Images

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Product Description

The undiluted product has been shown to be stable over several freeze/thaw cycles.

Product Specific Literature References

Molecular cloning and characterization of two novel pro-apoptotic isoforms of caspase-10: P.W. Ng, et al.; J. Biol. Chem. 274, 10301 (1999) Abstract
Caspase-8 and caspase-10 activate NF-kappaB through RIP, NIK and IKKalpha kinases: Y. Shikama, et al.; Eur. J. Immunol. 33, 1998 (2003) Abstract
Proteasome inhibition results in TRAIL sensitization of primary keratinocytes by removing the resistance-mediating block of effector caspase maturation: M. Leverkus, et al.; Mol. Cell Biol. 23, 777 (2003) Abstract
Caspase-10 is recruited to and activated at the native TRAIL and CD95 death-inducing signalling complexes in a FADD-dependent manner but can not functionally substitute caspase-8: M.R. Sprick, et al.; EMBO J. 21, 4520 (2002) Abstract

Further Categories Containing This Product:

Enzymes • Recombinant Proteins/Fusion Proteins

ALX-201-092

Revised 09-Aug-05

Caspase-10/b (active) (human) (recombinant)
PRODUCT LINE	Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY	Caspases

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-092-U025		25 U	203.00 USD
ALX-201-092-U100		100 U	371.00 USD

Product Specification

EC:	3.4.22.-
SOURCE/HOST:	Produced in E. coli.
PURITY:	≥90% (SDS-PAGE)
FORMULATION:	Lyophilized.
RECONSTITUTION:	Reconstitute with PBS to 1U/µl.
SPECIFIC ACTIVITY:	~15'000U/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.
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

In vitro activation of CPP32 and Mch3 by Mch4, a novel human apoptotic cysteine protease containing two FADD-like domains: T. Fernandes-Alnemri, et al.; PNAS 93, 7464 (1996) Abstract
Fas-associated death domain protein interleukin-1beta-converting enzyme 2 (FLICE2), an ICE/Ced-3 homologue, is proximally involved in CD95- and p55-mediated death signaling: C. Vincenz & V.M. Dixit; J. Biol. Chem. 272, 6578 (1997) Abstract; Full Text

General Information

BACKGROUND/TECHNICAL INFORMATION

Swiss-Prot link Q92851: Caspase-10 (human) (precursor)
AfCS Signalling Gateway link A000493: Caspase-10 (mouse)

Further Categories Containing This Product:

Enzymes • Recombinant Proteins/Fusion Proteins

ALX-201-194

Revised 06-Nov-07

Caspase-10/d (active) (human) (recombinant) (high stability)
PRODUCT LINE	Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY	Caspases

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-194-U050		50 U	320.00 USD

Product Specification

MW:	~22kDa (large subunit); ~12kDa (small subunit).
EC:	3.4.22.-
SOURCE/HOST:	Produced in E. coli.
CONCENTRATION:	~1mg/ml
PURITY:	≥95% (SDS-PAGE)
FORMULATION:	Liquid. 20µl in 100mM HEPES, pH 7.0, containing 10% sucrose, 10mM DTT, 2mM EDTA and 0.1% CHAPS.
SPECIFIC ACTIVITY:	~2'500U/mg protein. One unit is defined as the amount of enzyme that cleaves 1nmol of the caspase substrate IETD-pNA per min. at 37°C in a buffered solution.
SHIPPING:	SHIPPED ON DRY ICE
LONG TERM STORAGE:	-80°C

Product Images

Please click on thumbnails to enlarge.

Product Specific Literature References

Further Categories Containing This Product:

Enzymes • Recombinant Proteins/Fusion Proteins

ALX-850-244

Revised 22-Apr-08

CaspSCREEN™ Flow Cytometric Apoptosis Detection Kit
PRODUCT LINE	Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY	Caspase Kits & Sets

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-850-244-KI01		25 tests	203.00 USD
ALX-850-244-KI02		100 tests	469.00 USD

Product Specification

KIT/SET CONTAINS:	The kit includes D₂R reagent, DTT and the optimal D₂R incubation buffer together with an application protocol.
SHIPPING:	SHIPPED ON BLUE ICE
LONG TERM STORAGE:	-20°C
HANDLING:	Protect from light.

Product Description

The kit provides a convenient means for detecting activation of caspases by flow cytometry in living cells. The assay is based on the cleavage of (aspartyl)₂-rhodamine 110 (D₂R), a reported substrate for members of the caspase family proteases. The caspase substrate D₂R is non-fluorescent, however, upon cleavage of the substrate by cellular caspases, the released rhodamine 110 gives rise to fluorescence that can be measured at an excitation of 488nm and an emission of 530nm.

General Information

MANUFACTURER

Manufactured by BioVision.

Further Categories Containing This Product:

Detection Kits

ALX-260-094

Revised 09-May-06

D2R (high purity)