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Recombinant Human Caspase-7 Protein

Catalog #: 823-C7
6 Citations Datasheet / COA / SDS
Newer Version Available: 823-C7/CF
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Discontinued Product

823-C7 has been discontinued and is replaced by 823-C7/CF.

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Recombinant Human Caspase-7 Protein Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to cleave the fluorogenic peptide substrate Ac-DEVD-AFC. The specific activity is >3300 pmol/min/µg, as measured under the described conditions.
Source
E. coli-derived human Caspase-7 protein
Ala24-Asp198 (p20) & Ala207-Gln303 (p11)
Accession #
P55210
N-terminal Sequence
Analysis
Ala24 (p20) & Ala207 (p11)
Predicted Molecular Mass
20 kDa (p20) & 11 kDa (p11)
SDS-PAGE
20 kDa and 10 kDa, reducing conditions

Product Datasheets

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823-C7 (with carrier)

Product Datasheet
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823-C7/CF (carrier free)

Product Datasheet
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Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

823-C7

Formulation Supplied as a 0.2 μm filtered solution in HEPES, NaCl, DTT and Sucrose with BSA as a carrier protein.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

823-C7/CF

Formulation Supplied as a 0.2 μm filtered solution in HEPES, NaCl, DTT and Sucrose.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 25 mM HEPES, 0.1% (w/v) CHAPS, 10 mM dithiothreitol (DTT), pH 7.5
  • Recombinant Human Caspase-7 (rhCaspase-7) (Catalog # 823-C7)
  • Substrate: Ac-Asp-Glu-Val-Asp-AFC (MP Biomedicals, Catalog # AFC138), 10 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhCaspase-7 to 0.2 ng/µL in Assay Buffer.
  2. Dilute Substrate to 100 µM in Assay Buffer.
  3. Load 50 µL of 0.2 ng/µL rhCaspase-7 into a plate, and start the reaction by adding 50 µL of 100 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 100 µM Substrate.
  4. Read at excitation and emission wavelengths of 400 nm and 505 nm (top read), respectively, in kinetic mode for 5 minutes.
  5. Calculate specific activity:

         Specific Activity (pmol/min/µg) =

    		 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
    amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard 7-amino, 4-(trifluoromethyl)coumarin (Calbiochem, Catalog # 164580).

Per Well:
  • rhCaspase-7: 0.010 µg
  • Substrate: 50 µM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Caspase-7

Caspase-7 (Cysteine-aspartic acid protease 7/Casp7; also CMH-1, ICE-LAP3 and Mch3) is a 32 kDa member of the peptidase C14A/IL-1 beta -converting family of enzymes (1, 2, 3). It is widely expressed, except in brain, and is best known as an integral component of the apoptotic cascade. Caspase-7 is considered to be an executioner caspase, as a downstream mediator of apoptotic-associated proteolysis (2, 3). Upon activation, Caspase-7 is known to utilize a Cys residue to cleave multiple substrates, including PARP, procaspase 6, Gas2 and calpstatin (1). Human procaspase-7 is a 34-36 kDa, 303 amino acid (aa) protein (4, 5, 6). Normally, it is an inactive homodimer (1, 2, 7, 8). But following an upstream signal that activates processing proteases, procaspase-7 undergoes proteolytic cleavage to generate an N-terminal 23 aa propeptide, a 175 aa p20/20 kDa subunit (aa 24-198), and a 105 aa C-terminal p12/12 kDa subunit (5). The p20 and p12 subunits noncovalently heterodimerize, and subsequently associate with another p20/p12 heterodimer to form an active antiparallel homodimer. Additional processing of p20 may remove aa 24‑36 to generate p18, while additional processing of p12 will remove aa 199‑206 to generate p11 (9, 10). Multiple proteases can use Caspase-7 as a substrate, and include caspase-1, -3, -8, and -10, granzyme B, calpain-1 and Caspase-7 itself (3, 6, 9, 11). Caspase-7 is found in both cytosol and nucleus, and possesses a potential KKKK nuclear localization signal between aa 38-41 that likely undergoes sumoylation (9, 12). There are two potential isoform variants, one which shows an alternate start site 33 aa upstream of the standard start site, and a second that shows a 105 aa substitution for aa 149-303. Human and mouse Caspase-7 are 82% aa identical at the amino acid level.

References
  1. Chowdhury, I. et al. (2008) Comp. Biochem. Physiol. B 151:10.
  2. Boatright, K.M. and G.S. Salvesen (2003) Curr. Opin. Cell Biol. 15:725.
  3. Launay, S. et al. (2005) Oncogene 24:5137.
  4. Juan, T. et al. (1997) Genomics 40:86.
  5. Fernandez-Alnemri, T. et al. (1995) Cancer Res. 55:6045.
  6. Fernandez-Alnemri, T. et al. (1996) Proc. Natl. Acad. Sci. USA 93:7464.
  7. Gao, Z. et al. (2007) J. Biol. Chem. 282:30718.
  8. Riedl, S.J. et al. (2001) Proc. Natl. Acad. Sci. USA 98:14790.
  9. Gafni, J. et al. (2009) J. Biol. Chem. July 21 [epub ahead of print].
  10. Lippke, J.A. et al. (1996) J. Biol. Chem. 271:1825.
  11. Lamkanfi, M. et al. (2008) Mol. Cell. Proteomics 7:2350.
  12. Hayashi, N. et al. (2006) Neurosci. Lett. 397:5.
Entrez Gene IDs
840 (Human); 12369 (Mouse)
Alternate Names
apoptosis-related cysteine protease; Apoptotic protease Mch-3; CASP7; CASP-7; caspase 7, apoptosis-related cysteine peptidase; Caspase7; Caspase-7; EC 3.4.22; EC 3.4.22.60; ICE-like apoptotic protease 3; Lice2 alpha/beta/gamma; Mch3

Citations for Recombinant Human Caspase-7 Protein

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

6 Citations: Showing 1 - 6
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  1. Caspase-mediated cleavage of nucleocapsid protein of a protease-independent porcine epidemic diarrhea virus strain
    Authors: C Oh, Y Kim, KO Chang
    Virus Res., 2020-05-18;285(0):198026.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay
  2. Caspase-Mediated Cleavage of Nucleocapsid protein of a Protease-Independent Porcine Epidemic Diarrhea Virus Strain
    Authors: C Oh, Y Kim, KO Chang
    Virus Res., 2020-01-01;285(0):198026.
    Species: Human
    Sample Types: Plasmid
    Applications: Bioassay
  3. Fluorogenic Substrates for In Situ Monitoring of Caspase-3 Activity in Live Cells
    PLoS ONE, 2016-05-11;11(5):e0153209.
    Species: Human
    Sample Types: Peptide
    Applications: Bioassay
  4. Anamorsin, a novel caspase-3 substrate in neurodegeneration.
    Authors: Yun N, Lee Y, Kim C, Shibayama H, Tanimura A, Hamanaka Y, Kanakura Y, Park I, Jo A, Shin J, Ju C, Kim W, Oh Y
    J Biol Chem, 2014-06-27;289(32):22183-95.
    Species: Mouse
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  5. Degradomics reveals that cleavage specificity profiles of caspase-2 and effector caspases are alike.
    Authors: Wejda M, Impens F, Takahashi N, Van Damme P, Gevaert K, Vandenabeele P
    J Biol Chem, 2012-07-23;287(41):33983-95.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Enzyme Assay
  6. Fine-tuning nucleophosmin in macrophage differentiation and activation.
    Authors: Guery L, Benikhlef N, Gautier T, Paul C, Jego G, Dufour E, Jacquel A, Cally R, Manoury B, Vanden Berghe T, Vandenabeele P, Droin N, Solary E
    Blood, 2011-08-29;118(17):4694-704.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay

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