Recombinant Human Pro-Caspase-3 Protein, CF

R&D Systems, part of Bio-Techne | Catalog # 731-C3

R&D Systems, part of Bio-Techne
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731-C3-005
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Key Product Details

Source

E. coli

Accession #

AAA65015

Structure / Form

Pro form

Conjugate

Unconjugated

Applications

Enzyme Activity

View all Caspase-3 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Caspase-3 protein
Met1-His277, with an N-terminal Met and 6-His tag

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

N-terminal Sequence Analysis

Not tested: Met predicted.

Predicted Molecular Mass

33 kDa

SDS-PAGE

35 kDa, reducing conditions

Activity

The activity of the activated Caspase-3 was measured by its ability to cleave DEVD-AFC.
The specific activity is >500 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation and Storage

731-C3
Formulation Lyophilized from a 0.2 μm filtered solution in Tris, NaCl, Sucrose and DTT.
Reconstitution Reconstitute at 50 µg/mL in 20 mM Tris, 300 mM NaCl, 5 mM Dithiothreitol, 5% Sucrose and 0.05% CHAPS, pH 8.0.
Shipping The product is shipped at ambient temperature. 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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

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

UniProt

AAA65015

Additional Caspase-3 Products

Product Documents for Recombinant Human Pro-Caspase-3 Protein, CF

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

For research use only

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