218720 Caspase-4, Human, Recombinant, E. coli

218720
  
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      Přehled

      Replacement Information

      Tabulka spec. kláve

      Description
      Overview

      This product has been discontinued.





      Recombinant, human caspase-4 expressed in E. coli. Suitable for the study of enzyme regulation and kinetics, target substrate cleavage, and inhibitor screening. MW: 10000 and 19000.
      M.W. 10,000 and 19,000 (heterodimer).
      Catalogue Number218720
      Brand Family Calbiochem®
      SynonymsICErel-II, ICH-2, TX
      References
      ReferencesKamada, S., et al. 1997. Oncogene, 15, 285.
      Nasir, J., et al. 1997. Mamm. Genome 8, 611.
      Nath, R., et al. 1996. Biochem. J. 319, 683.
      Product Information
      Unit of DefinitionOne unit is defined as the amount of enzyme that will cleave 1.0 pmol substrate Ac-LEHD-<i>p</i>NA (Cat. No. 218805) per min at 25°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.
      Applications
      Biological Information
      Purity≥90% by SDS-PAGE
      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-4, Human, Recombinant, E. coli Certificates of Analysis

      TitleLot Number
      218720

      References

      Přehled odkazů
      Kamada, S., et al. 1997. Oncogene, 15, 285.
      Nasir, J., et al. 1997. Mamm. Genome 8, 611.
      Nath, R., et al. 1996. Biochem. J. 319, 683.

      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.

      Revision31-January-2008 JSW
      SynonymsICErel-II, ICH-2, TX
      DescriptionRecombinant, human caspase-4 expressed in E. coli. Suitable for the study of enzyme regulation and kinetics, target substrate cleavage, and inhibitor screening. Member of the caspase-1 or ICE (interleukin-1β-converting enzyme) subfamily that is involved in Fas-mediated apoptosis. Cleavage sites of the maturation process of caspase-4 are located at Asp104, Asp270, and Asp289. Interacts with Apaf-1 and participates in the cytochrome c activation of caspase-3. Also associates with caspase-14, a short prodomain caspase. MW: 10000 and 19000.
      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
      Activity Assay Required Materials • Assay Buffer: 100 mM NaCl, 50 mM HEPES, 10 mM DTT, 1 mM EDTA, 10% glycerol, 0.1% CHAPS • Substrate: 100 mM stock of appropriate substrate (e.g. Ac-LEHD-pNA, Cat. No. 218805), prepared in DMSO; diluted to 400 µM in Assay Buffer just prior to use (final concentration in the well = 200 µM) • Caspase-4: dilute to 2 U/µl just prior to use (final amount of enzyme per well = 30 U) • Spectrophotometer: capacity to measure at 405 nm • 1/2 Volume 96-Well Plate
      Protocol: 1. Add 35 µl Assay Buffer to the desired number of wells in a 1/2 Volume 96-Well Plate; allow to equilibrate to 25°C. 2. Add 15 µl Caspase-4 (2 U/µl) to each well; include 2 blanks that contain 15 µl Assay Buffer in place of the enzyme. 3. To start the reaction, add 50 µl Substrate (400 µM in assay buffer); mix by pipetting up and down. 4. Continuously monitor the absorbance at 405 nm. 5. Data analysis: plot absorbance vs. time and determine the slope over the linear portion of the curve; convert change in absorbance/minute to substrate/minute using an extinction coefficient of 10,500 M-1cm-1 for pNA. Correct the values for a light path of ~0.5 cm and a 100 µl well of a 1/2 Volume Plate.
      Purity≥90% by SDS-PAGE
      Unit definitionOne unit is defined as the amount of enzyme that will cleave 1.0 pmol substrate Ac-LEHD-pNA (Cat. No. 218805) per min at 25°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
      ReferencesKamada, S., et al. 1997. Oncogene, 15, 285.
      Nasir, J., et al. 1997. Mamm. Genome 8, 611.
      Nath, R., et al. 1996. Biochem. J. 319, 683.