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Recombinant Human Caspase-3 Protein, CF

Catalog #: 707-C3/CF
16 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
707-C3-010/CF
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Citations (16)
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Recombinant Human Caspase-3 Protein, CF Summary

Product Specifications

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.
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.
Source
E. coli-derived human Caspase-3 protein
Ser29-Asp175 (subunit 1) & Ala183-His277 (Asp190Glu) (subunit 2)
Accession #
P42574
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

Product Datasheets

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707-C3/CF (carrier free)

Product Datasheet
COA
SDS

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.

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.

Assay Procedure

Materials
  • Assay Buffer: 25 mM HEPES, 0.1% (w/v) CHAPS, 10 mM dithiothreitol (DTT), pH 7.5
  • Recombinant Human Caspase-3 (rhCaspase-3) (Catalog # 707-C3/CF)
  • 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 3 to 0.4 ng/µL in Assay Buffer.
  2. Dilute Substrate to 100 µM in Assay Buffer.
  3. Load 50 µL of 0.4 ng/µL rhCaspase 3 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-3: 0.02 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-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 & -δ, 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.
Entrez Gene IDs
836 (Human); 12367 (Mouse)
Alternate Names
Apopain; apoptosis-related cysteine protease; CASP3; CASP-3; caspase 3, apoptosis-related cysteine peptidase; Caspase3; Caspase-3; CPP32; CPP-32; CPP32B; CPP32SREBP cleavage activity 1; Cysteine protease CPP32; EC 3.4.22; EC 3.4.22.56; LICE-1; PARP cleavage protease; procaspase3; Protein Yama; SCA-1; YAMA

Citations for Recombinant Human Caspase-3 Protein, CF

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.

16 Citations: Showing 1 - 10
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  1. Mannose antagonizes GSDME-mediated pyroptosis through AMPK activated by metabolite GlcNAc-6P
    Authors: Ai, YL;Wang, WJ;Liu, FJ;Fang, W;Chen, HZ;Wu, LZ;Hong, X;Zhu, Y;Zhang, CX;Liu, LY;Hong, WB;Zhou, B;Chen, QT;Wu, Q;
    Cell research
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  2. Genuine selective caspase-2 inhibition with new irreversible small peptidomimetics
    Authors: E Bosc, J Anastasie, F Soualmia, P Coric, JY Kim, LQ Wang, G Lacin, K Zhao, R Patel, E Duplus, P Tixador, AA Sproul, B Brugg, M Reboud-Rav, CM Troy, ML Shelanski, S Bouaziz, M Karin, C El Amri, ED Jacotot
    Cell Death & Disease, 2022-11-15;13(11):959.
    Species: N/A
    Sample Types: Fluorogenic Peptide Substrate
    Applications: Bioassay
  3. The metabolite &alpha-KG induces GSDMC-dependent pyroptosis through death receptor 6-activated caspase-8
    Authors: JY Zhang, B Zhou, RY Sun, YL Ai, K Cheng, FN Li, BR Wang, FJ Liu, ZH Jiang, WJ Wang, D Zhou, HZ Chen, Q Wu
    Cell Research, 2021-05-19;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  4. Autophagy controls the induction and developmental decline of NMDAR-LTD through endocytic recycling
    Authors: H Shen, H Zhu, D Panja, Q Gu, Z Li
    Nat Commun, 2020-06-12;11(1):2979.
    Species: Mouse
    Sample Types: Organotypic Tissue Slice
    Applications: Bioassay
  5. Eradication of unresectable liver metastasis through induction of tumour specific energy depletion
    Authors: D Huo, J Zhu, G Chen, Q Chen, C Zhang, X Luo, W Jiang, X Jiang, Z Gu, Y Hu
    Nat Commun, 2019-07-11;10(1):3051.
    Species: Human
    Sample Types: Nanoparticles
    Applications: Bioassay
  6. Cilostazol protects hepatocytes against alcohol-induced apoptosis via activation of AMPK pathway
    Authors: YJ Lee, MS Shu, JY Kim, YH Kim, KH Sim, WJ Sung, JR Eun
    PLoS ONE, 2019-01-29;14(1):e0211415.
    Applications: Bioassay
  7. Tom20 senses iron-activated ROS signaling to promote melanoma cell pyroptosis
    Authors: B Zhou, JY Zhang, XS Liu, HZ Chen, YL Ai, K Cheng, RY Sun, D Zhou, J Han, Q Wu
    Cell Res., 2018-10-04;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  8. Alkyne-based surface-enhanced Raman scattering nanoprobe for ratiometric imaging analysis of caspase 3 in live cells and tissues
    Authors: X Qin, M Lyu, Y Si, J Yang, Z Wu, J Li
    Anal. Chim. Acta, 2018-09-08;1043(0):115-122.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  9. Active site-targeted covalent irreversible inhibitors of USP7 impair the functions of Foxp3+ T-regulatory cells by promoting ubiquitination of Tip60
    Authors: F Wang, L Wang, J Wu, I Sokirniy, P Nguyen, T Bregnard, J Weinstock, M Mattern, I Bezsonova, WW Hancock, S Kumar
    PLoS ONE, 2017-12-13;12(12):e0189744.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  10. Caspase-3/-7-Specific Metabolic Precursor for Bioorthogonal Tracking of Tumor Apoptosis
    Authors: MK Shim, HY Yoon, S Lee, MK Jo, J Park, JH Kim, SY Jeong, IC Kwon, K Kim
    Sci Rep, 2017-11-30;7(1):16635.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  11. 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
  12. 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
  13. In vivo and in vitro models for the therapeutic targeting of Wnt signaling using a Tet-ODeltaN89beta-catenin system.
    Authors: Jarde T, Evans R, McQuillan K, Parry L, Feng G, Alvares B, Clarke A, Dale T
    Oncogene, 2012-04-02;32(7):883-93.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-P
  14. 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
  15. Interactions between important regulatory proteins and human alphaB crystallin.
    Authors: Ghosh JG, Shenoy AK, Clark JI
    Biochemistry, 2007-05-08;46(21):6308-17.
    Species: Human
    Sample Types: Peptide
    Applications: ELISA-Based Protein Pin Array
  16. Acquired resistance to TRAIL-induced apoptosis in human ovarian cancer cells is conferred by increased turnover of mature caspase-3.
    Authors: Lane D, Cote M, Grondin R, Couture MC, Piche A
    Mol. Cancer Ther., 2006-03-01;5(3):509-21.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Enzyme Assay

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