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235417 Caspase-3, Human, Recombinant, E. coli

235417
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

Products

Catalogue NumberPackaging Qty/Pack
235417-5000U Plastic ampoule 5000 u
Description
OverviewRecombinant, human caspase-3 expressed in E. coli as the proenzyme, which is subsequently cleaved to produce the active enzyme. Suitable for the study of enzyme regulation, kinetics, and target substrate cleavage and inhibitor screening. MW: 12000 and 17000.
M.W. 12,000 and 17,000 (heterodimer).
Catalogue Number235417
Brand Family Calbiochem®
SynonymsYAMA, Apopain, CPP32
References
ReferencesIzban, K.F., et al. 1999. Am. J. Pathol. 154, 1439.
Thornberry, N.A., and Lazebnik, Y. 1998. Science 281, 1312.
Mittl, P.R.E., et al. 1997. J. Biol. Chem. 272, 6539.
Talanian, R.V., et al. 1997. J. Biol. Chem. 272, 9677.
Thornberry, N.A., et al. 1997. J. Biol. Chem. 272, 17907.
Product Information
Unit of DefinitionOne unit is defined as the amount of enzyme that will cleave 1.0 pmol of the substrate Ac-DEVD-<i>p</i>NA (<a href ="/Products/ProductDisplay.asp→catNO=235400">Cat. No. 235400</a>) per min at 30°C, pH 7.4.
FormLiquid
FormulationIn 100 mM NaCl, 50 mM HEPES, 10 mM DTT, 1 mM EDTA, 10% glycerol, 0.5% CHAPS, pH 7.4.
Quality LevelMQ100
Applications
Biological Information
Purity≥95% by SDS-PAGE
Specific Activity≥1450units/µg protein
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Storage and Shipping Information
Ship Code Dry Ice Only
Toxicity Standard Handling
Storage ≤ -70°C
Avoid freeze/thaw Avoid freeze/thaw
Do not freeze Ok to freeze
Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
Packaging Information
Transport Information
Supplemental Information
Specifications

Documentation

Caspase-3, Human, Recombinant, E. coli SDS

Title

Safety Data Sheet (SDS) 

Caspase-3, Human, Recombinant, E. coli Certificates of Analysis

TitleLot Number
235417

References

Reference overview
Izban, K.F., et al. 1999. Am. J. Pathol. 154, 1439.
Thornberry, N.A., and Lazebnik, Y. 1998. Science 281, 1312.
Mittl, P.R.E., et al. 1997. J. Biol. Chem. 272, 6539.
Talanian, R.V., et al. 1997. J. Biol. Chem. 272, 9677.
Thornberry, N.A., et al. 1997. J. Biol. Chem. 272, 17907.

Brochure

Title
Caspases and other Apoptosis Related Tools Brochure
Data Sheet

Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

Revision15-June-2018 JSW
SynonymsYAMA, Apopain, CPP32
DescriptionRecombinant, human caspase-3 expressed in E. coli as the proenzyme, which is subsequently cleaved to produce the active enzyme. The active enzyme is a heterodimer with subunit molecular weights of 12 kDa and 19 kDa. Suitable for the study of enzyme regulation and kinetics, target substrate cleavage and inhibitor screening. Caspase-3 is a member of the ICE family of cysteine proteases that is activated during apoptotic signaling events by upstream proteases, including caspase-6, caspase-8 (FLICE) and cytotoxic T-cell-derived granzyme B.
FormLiquid
FormulationIn 100 mM NaCl, 50 mM HEPES, 10 mM DTT, 1 mM EDTA, 10% glycerol, 0.5% CHAPS, pH 7.4.
Recommended reaction conditions
Caspase-3 Activity Assay This protocol is provided as a general guide; conditions should be optimized for individual experiments.
Materials Required •Assay Buffer: 100 mM NaCl, 50 mM HEPES, 10 mM DTT, 1 mM EDTA, 10% glycerol, 0.1% CHAPS, pH 7.4 •Caspase-3: Dilute the recombinant caspase-3 to 10 U/µl in Assay Buffer just prior to use. •Caspase-3 Substrate I, Colorimetric (Cat. No. 235400): Prepare a 20 mM stock solution in DMSO (for 5 mg peptide add 392 µl DMSO). Dilute 1:10 in Assay Buffer (2 mM final concentration) just prior to use. •Half-volume 96-well plate Protocol: Reaction Conditions at Vmax (saturated substrate):
1. Add 87 µl Assay Buffer into a half-volume 96-well plate. Allow the plate to equilibrate to room temperature (20°C). 2. Add 3 µl Caspase-3 (10 U/µl) to each well (leave 2 blanks that do not contain Caspase-3). 3. To start the reaction, add 10 µl Caspase-3 Substrate I, Colorimetric (2 mM in assay buffer). The final substrate concentration per well is 200 µM. 4. Continuously monitor the absorbance at 405nm. 5. Graph absorbance vs. time and determine the slope during from the linear part of the curve. Convert to substrate amounts using the extinction coefficient for p-nitroaniline (~10,000 M-1cm-1) and adjusting for pathlength.
Purity≥95% by SDS-PAGE
Specific activity≥1450units/µg protein
Unit definitionOne unit is defined as the amount of enzyme that will cleave 1.0 pmol of the substrate Ac-DEVD-pNA (Cat. No. 235400) per min at 30°C, pH 7.4.
Storage ≤ -70°C
Avoid freeze/thaw
Do Not Freeze Ok to freeze
Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
Toxicity Standard Handling
ReferencesIzban, K.F., et al. 1999. Am. J. Pathol. 154, 1439.
Thornberry, N.A., and Lazebnik, Y. 1998. Science 281, 1312.
Mittl, P.R.E., et al. 1997. J. Biol. Chem. 272, 6539.
Talanian, R.V., et al. 1997. J. Biol. Chem. 272, 9677.
Thornberry, N.A., et al. 1997. J. Biol. Chem. 272, 17907.