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ALX-201-038
Revised 24-Jan-05
Caspase-3 (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-201-038-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
[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-3 spontaneously undergoes autoprocessing to yield the subunits characteristic of the active enzyme.
The rate of caspase-3 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 P42574:
Caspase-3 (human) (precursor)
AfCS Signalling Gateway link A000498:
Caspase-3 (mouse)
Further Categories Containing This Product:
Enzymes
•
Recombinant Proteins / Fusion Proteins
ALX-201-059
Revised 21-Jul-08
Caspase-3 (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-201-059-U025
25 U
119.00 USD
ALX-201-059-U100
100 U
273.00 USD
Product Specification
EC:
3.4.22.56
SOURCE/HOST:
Produced in
E. coli
.
FORMULATION:
Lyophilized.
RECONSTITUTION:
Reconstitute to 1U/µl with PBS containing 15% glycerol.
SPECIFIC ACTIVITY:
300’000-400’000U/mg protein. One unit is defined as the amount of enzyme that cleaves 1nmol of the caspase substrate DEVD-
p
NA (Prod. No.
ALX-260-033
) 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 studying enzyme regulation, determining target substrate, screening caspase inhibitors, or as a positive control in caspase assays. We recommend using 1 unit per assay for analyzing caspase activity.
For a complete caspase-3 assay protocol, please refer to Caspase-3 Fluorometric or Colorimetric Assay Kits (Prod. No.
ALX-850-216
or
ALX-850-215
).
QUALITY CONTROL:
Routinely tested for its ability to cleave the caspase-3 substrates Ac-DEVD-
p
NA (Prod. No.
ALX-260-033
) and Ac-DEVD-AFC (Prod. No.
ALX-260-032
).
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-3 is a member of the interleukin-1β converting enzyme (ICE) family of cysteine proteases. It exists in cells as an inactive 32kDa proenzyme. During apoptosis procaspase-3 is processed at aspartate residues by self-proteolysis and/or cleavage by upstream caspases, such as caspase-6, caspase-8 and granzyme B. The processed form of caspase-3 consists of large (17kDa) and small (11kDa) subunits which associate to form the active enzyme. Active caspase-3 has been shown involved in the proteolysis of several important molecules, such as poly(ADP-ribose) polymerase (PARP), the sterol regulatory element binding proteins (SREBPs), focal adhesion kinase (FAK) and others.
BACKGROUND/TECHNICAL INFORMATION
Swiss Prot link:
Caspase-3 (human) (precursor)
AfCS Signalling Gateway link:
Caspase-3 (mouse)
Further Categories Containing This Product:
Enzymes
•
Recombinant Proteins / Fusion Proteins
ALX-522-069
Revised 30-Jun-05
Caspase-3 (active) (human) (recombinant) (high purity)
PRODUCT LINE
Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY
Caspases
Ordering Information
Product Numbers:
Format:
Size:
Unit Price:
Quantity:
Add To Cart
ALX-522-069-C005
5 µg
180.00 USD
Product Specification
MW:
~29kDa; α-subunit: ~17kDa; β-subunit: ~12kDa.
EC:
3.4.22.-
SOURCE/HOST:
Produced in
E. coli.
CONCENTRATION:
0.1mg/ml
PURITY:
≥95% (SDS-PAGE)
FORMULATION:
Liquid. In 20mM PIPES, pH 7.4, containing 10mM DTT, 100mM sodium chloride, 10% (w/v) sucrose, 0.1% (w/v) CHAPS and 1mM EDTA.
SPECIFIC ACTIVITY:
≥50'000U/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 min. at 37°C in a buffered solution.
SHIPPING:
SHIPPED ON DRY ICE
LONG TERM STORAGE:
-80°C
HANDLING:
Avoid freeze/thaw cycles.
Product Images
Please click on thumbnails to enlarge.
General Information
BACKGROUND/TECHNICAL INFORMATION
The rate of recombinant human caspase-3 enzymatic activity can be measured by the release of AMC from the Ac-DEVD-AMC (Prod. No.
ALX-260-031
) caspase substrate as emission at 440nm upon excitation at 380nm using a spectrofluorometer [1].
Swiss-Prot link:
Caspase-3 (human) (precursor)
AfCS Signalling Gateway link:
Caspase-3 (mouse)
MANUFACTURER
Manufactured by Apotech Corporation.
General 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
Further Categories Containing This Product:
Enzymes
•
Recombinant Proteins / Fusion Proteins
ALX-201-162
Revised 08-Dec-04
Caspase-3 (active) (mouse) (recombinant)
PRODUCT LINE
Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY
Caspases
Ordering Information
Product Numbers:
Format:
Size:
Unit Price:
Quantity:
Add To Cart
ALX-201-162-C050
50 µg
280.00 USD
Product Specification
SOURCE/HOST:
Produced in
E. coli.
CONCENTRATION:
0.81mg/ml
PURITY:
~95% (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:
~1'500U/µg protein. One unit is defined as the amount of enzyme that cleaves 1nmol of the caspase substrate Z-DEVD-AMC (AMC: 7-aminomethylcoumarin) per hour at 30°C in a reaction solution containing 20µM Z-DEVD-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.
Further Categories Containing This Product:
Enzymes
•
Recombinant Proteins / Fusion Proteins
ALX-201-078
Revised 24-Jan-05
Caspase-3 (rat) (recombinant)
PRODUCT LINE
Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY
Caspases
Ordering Information
Product Numbers:
Format:
Size:
Unit Price:
Quantity:
Add To Cart
ALX-201-078-C005
5 µg
320.00 USD
Product Specification
EC:
3.4.22.-
SOURCE/HOST:
Produced in
E. coli
.
CONCENTRATION:
0.165mg/ml
PURITY:
≥95% (SDS-PAGE)
FORMULATION:
Liquid. 5µg of caspase-3 in 50mM HEPES, pH 7.4, 100mM NaCI, 0.5% CHAPS, 10mM DTT, 1mM EDTA and 50% glycerol.
SPECIFIC ACTIVITY:
≥
500U
/µg protein. One unit is defined as the amount of enzyme that cleaves 1nmol of the caspase-substrate Z-DEVD-AMC per hour at 20°C in a reaction solution containing 20µM substrate, 50mM HEPES, pH 7.4, 100mM NaCl, 0.5% CHAPS, 10mM DTT, 1mM EDTA and 10% glycerol.
APPLICATION:
In combination with caspase-3 activity assays, this product is useful in biological screening of caspase substrates and inhibitors. It can also be used as a positive control in caspase-3 assays as well as a positive control in Western blot.
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.
Product Description
Contains both the proenzyme and the processed (active) form.
Product Specific Literature References
Presence of DNA fragmentation and lack of neuroprotective effect in DFF45 knockout mice subjected to traumatic brain injury
:
A.G. Yakovlev, et al.; Mol. Med.
7
, 205 (2001)
Abstract
General Information
BACKGROUND/TECHNICAL INFORMATION
Enzyme Evaluation Protocol (general)
This protocol can be used to measure caspase enzyme activity. A synthetic fluorogenic peptide, e. g. Ac-DEVD-AMC (Prod. No.
ALX-260-031
) is used as the caspase enzyme substrate. The enzyme cleaves the substrate releasing the fluorescent AMC. AMC release can be measured by spectrofluorometry using UV.
1. Add 20µM of Ac-DEVD-AMC to 1 ml of assay buffer (20mM PIPES, pH 7.2, 100mM sodium chloride, 10mM DTT, 1mM EDTA, 0.1% (w/v) CHAPS, 10% sucrose). Add the appropriate amount of active caspase to the mixture (e.g. 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.
Swiss-Prot link P55213:
Caspase-3 (rat)
AfCS Signalling Gateway link A000498:
Caspase-3 (mouse)
Further Categories Containing This Product:
Enzymes
•
Recombinant Proteins / Fusion Proteins
ALX-850-215
Revised 22-Apr-08
Caspase-3 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-850-215-KI01
25 tests
175.00 USD
Product Specification
KIT/SET CONTAINS:
25ml Cell Lysis Buffer, 2ml 2X Reaction Buffer, 125µl DEVD-pNA (4mM), 100µl
DTT (1M), 25ml Dilution Buffer.
SHIPPING:
SHIPPED ON BLUE ICE
LONG TERM STORAGE:
-20°C
HANDLING:
Protect from light. Store Lysis Buffer, 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 DEVD-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-3 activity.
MANUFACTURER
Manufactured by BioVision.
Further Categories Containing This Product:
Activity Assays
ALX-151-031
Revised 23-Nov-06
Caspase-3 Control Peptide
PRODUCT LINE
Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY
Caspase Inhibitors
Ordering Information
Product Numbers:
Format:
Size:
Unit Price:
Quantity:
Add To Cart
ALX-151-031-M001
1 mg
35.00 USD
ALX-151-031-M005
5 mg
120.00 USD
Product Specification
SEQUENCE:
Ac-Asp-Glu-Val-Asp-OH
FORMULA:
C
20
H
30
N
4
O
12
MW:
518.5
PURITY:
≥97%
APPEARANCE:
Lyophilized.
SOLUBILITY:
Soluble in water.
SHIPPING:
AMBIENT
LONG TERM STORAGE:
-20°C
Further Categories Containing This Product:
Peptides
ALX-850-216
Revised 24-Jun-08
Caspase-3 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-850-216-KI01
25 tests
175.00 USD
Product Specification
KIT/SET CONTAINS:
25ml Cell Lysis Buffer, 2ml 2X Reaction Buffer, 125µl DEVD-AFC (1mM), 100µl DTT (1M).
SHIPPING:
SHIPPED ON BLUE ICE
LONG TERM STORAGE:
-20°C
HANDLING:
Protect from light. Store 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 DEVD-AFC.
Product Specific Literature References
Apoptosis and Id2 expression in diaphragm and soleus muscle from the emphysematous hamster:
H. Degens, et al.; Am. J. Physiol. Regul. Integr. Comp. Physiol.
293
, R135 (2007)
Abstract
General Information
DEVD-AFC emits blue light (λmax = 400nm); upon cleavage of the substrate by caspase-3 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-3 activity.
MANUFACTURER
Manufactured by BioVision.
Further Categories Containing This Product:
Activity Assays
ALX-201-093
Revised 15-Apr-08
Caspase-4 (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-201-093-U025
25 U
203.00 USD
ALX-201-093-U100
100 U
371.00 USD
Product Specification
EC:
3.4.22.57
SOURCE/HOST:
Produced in
E. coli
.
PURITY:
≥95% (SDS-PAGE)
FORMULATION:
Lyophilized.
RECONSTITUTION:
Reconstitute with PBS containing 15% glycerol to 1U/µl.
SPECIFIC ACTIVITY:
~5'000U/mg protein. One unit is defined as the amount of enzyme that cleaves 1nmol of the caspase substrate WEHD-pNA per hour at 37°C in 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 and kinetics, determining target substrate or as a 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
Caspase-4 is a member of the caspase family of cysteine proteases. It exists in cells as an inactive pro-enzyme. The pro-enzyme is matured by proteolysis to yield large and small subunits. The active enzyme is a heterotetramer consisting of two large and two small subunits. The expressed caspase-4 spontaneously undergoes auto-processing to yield subunits characteristic of the native enzyme. The active caspase-4 is routinely tested for its ability to enzymatically cleave the substrate WEHD-AFC or WEHD-pNA.
BACKGROUND/TECHNICAL INFORMATION
Swiss-Prot link P49662:
Caspase-4 (human) (precursor)
AfCS Signalling Gateway link A000499:
Caspase-4 (mouse)
Further Categories Containing This Product:
Enzymes
•
Endoplasmatic Reticulum Stress
•
Recombinant Proteins / Fusion Proteins
ALX-201-094
Revised 03-May-06
Caspase-5 (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-201-094-U025
25 U
203.00 USD
ALX-201-094-U100
100 U
371.00 USD
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 WEHD-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:
In combination with caspase-5 activity assays, the enzyme is useful in screening caspase inhibitors, studying enzyme regulation and kinetics, determining target substrate or as a positive control in caspase-5 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 P51878:
Caspase-5 (human) (precursor)
AfCS Signalling Gateway link A000500:
Caspase-5 (mouse)
Further Categories Containing This Product:
Inflammasome
•
Enzymes
•
Recombinant Proteins / Fusion Proteins
ALX-850-217
Revised 24-Jan-05
Caspase-5 Colorimetric Assay Kit
PRODUCT LINE
Cell Death / Apoptosis / Autophagy
