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

Caspase-3, Human, Recombinant, is expressed as a proenzyme in E. coli and is subsequently cleaved to produce the active enzyme.

Ficha de datos de seguridad (MSDS o SDS), certificado de análisis y de calidad (CoA y CoQ), expedientes, folletos y otros documentos disponibles.

Sinónimos: YAMA, Apopain, CPP32

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      Descripción

      Replacement Information
      Description
      Overview

      This product has been discontinued.



      Recombinant, 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≥1000 units/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
      Global Trade Item Number
      Número de referencia GTIN
      235417 0

      Documentation

      Caspase-3, Human, Recombinant, E. coli Ficha datos de seguridad (MSDS)

      Título

      Ficha técnica de seguridad del material (MSDS) 

      Caspase-3, Human, Recombinant, E. coli Certificados de análisis

      CargoNúmero de lote
      235417

      Referencias bibliográficas

      Visión general referencias
      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.

      Folleto

      Cargo
      Caspases and other Apoptosis Related Tools Brochure
      Ficha técnica

      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.

      Revision03-August-2020 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≥1000 units/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 Avoid freeze/thaw
      ≤ -70°C
      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.