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

235417
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      235417-5000UCN
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          Plastic ampoule 5000 u
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          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, CPP32, Apopain
          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 MSDS

          타이틀

          물질안전보건자료(MSDS) 

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

          TitleLot Number
          235417

          References

          참고문헌 보기
          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, CPP32, Apopain
          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.