Recombinant Human Caspase-3 Protein, CF

R&D Systems, part of Bio-Techne | Catalog # 707-C3/CF

R&D Systems, part of Bio-Techne
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707-C3-010/CF
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Conjugate
Catalog #
Unconjugated
707-C3
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Key Product Details

Source

E. coli

Conjugate

Unconjugated

Applications

Enzyme Activity

View all Caspase-3 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Caspase-3 protein
Ser29-Asp175 (subunit 1) & Ala183-His277 (Asp190Glu) (subunit 2)

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ser29 (subunit 1) & Ala183 (subunit 2)

Predicted Molecular Mass

17 kDa (subunit 1), 11 kDa (subunit 2)

SDS-PAGE

18 kDa and 10 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate Ac-DEVD-AFC.
The specific activity is >3,000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation and Storage

707-C3/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.

Background: Caspase-3

Caspase-3 (Cysteine-aspartic acid protease 3/Casp3; also Yama, apopain and CPP32) is a 29 kDa member of the peptidase C14A family of enzymes (1, 2, 3). It is widely expressed and is an integral component of the apoptotic cascade. Caspase-3 is considered to be the major executioner caspase; that is, the primary downstream mediator of apoptotic-associated proteolysis (2, 3, 4). Active Caspase-3 is known to utilize a Cys residue to cleave multiple substrates, including PARP, proIL‑16, PKC-gamma & -delta, procaspases 6, 7 and 9, and beta‑catenin (1). Human procaspase-3 is a 32 kDa, 277 amino acid (aa) protein (5, 6, 7). Normally, it is an inactive, cytosolic homodimer, but following an upstream signal that activates processing proteases, procaspase-3 undergoes proteolytic cleavage (1, 2, 8, 9). This generates an N-terminal 175 aa p20/20 kDa subunit plus a 102 aa C-terminal p12/12 kDa subunit, followed by further processing of the p20 subunit at Asp28 to generate a final p17 subunit (aa 29‑175) (9). The p17 and p12 subunits noncovalently heterodimerize, and subsequently associate with another p17/p12 heterodimer to form an active antiparallel homodimer. The p17 subunit contains the enzyme active site (aa 161‑165), with an embedded catalytic Cys which is normally nitrosylated and inactive. Full activation requires both proteolytic processing and Cys163 denitrosylation (10). Multiple proteases can use Caspase-3 as a substrate including Caspase-6, -8, and -10, granzyme B, and Caspase-3 itself (9, 11, 12, 13). 

References

  1. Chowdhury, I. et al. (2008) Comp. Biochem. Physiol. B 151:10.

  2. Boatright, K.M. & G.S. Salvesen (2003) Curr. Opin. Cell Biol. 15:725.

  3. Launay, S. et al. (2005) Oncogene 24:5137.

  4. Walsh, J.G. et al. (2008) Proc. Natl. Scad. Sci. USA 105:12815.

  5. Nicholson, D.W. et al. (1995) Nature 376:37.

  6. Tewari, M. et al. (1995) Cell 81:801.

  7. Fernandes-Alnemri, T. et al. (1994) J. Biol. Chem. 269:30761.

  8. Milisav, I. et al. (2009) Apoptosis 14:1070.

  9. Han, Z. et al. (1997) J. Biol. Chem. 272:13432.

  10. Rossig, L. et al. (1999) J. Biol. Chem. 274:6823.

  11. Rank, K.B. et al. (2001) Protein Expr. Purif. 22:258.

  12. Atkinson, E.A. et al. (1998) J. Biol. Chem. 273:21261.

  13. Cohen, G.M. (1997) Biochem. J. 326:1.

Alternate Names

Apopain, CASP3, Caspase3, CPP32, LICE-1, YAMA

Entrez Gene IDs

836 (Human); 12367 (Mouse)

Gene Symbol

CASP3

Additional Caspase-3 Products

Product Documents for Recombinant Human Caspase-3 Protein, CF

Product Datasheets

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Product Specific Notices for Recombinant Human Caspase-3 Protein, CF

For research use only

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