Recombinant Human Caspase-3 Protein

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Summary
Product Discontinued
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    • Catalog Number
      707-C3
    • Availability
      Product Discontinued

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Recombinant Human Caspase-3 Protein Summary

Details of Functionality
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 #
N-terminal Sequence
Ser29 (subunit 1) & Ala183 (subunit 2)
Protein/Peptide Type
Recombinant Enzymes
Gene
CASP3
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
17 kDa (subunit 1), 11 kDa (subunit 2).
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
18 kDa and 10 kDa, reducing conditions
Publications
Read Publications using
707-C3 in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in HEPES, NaCl, DTT and Sucrose with BSA as a carrier protein.
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • 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)
  • 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Caspase-3 Protein

  • 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

Background

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). 
  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.

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707-C3
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Publications for Caspase-3 (707-C3)(16)

We have publications tested in 3 confirmed species: Human, Mouse, N/A.

We have publications tested in 4 applications: Bioassay, ELISA-Based Protein Pin Array, Enzyme Assay, IHC-P.


Filter By Application
Bioassay
(10)
ELISA-Based Protein Pin Array
(1)
Enzyme Assay
(4)
IHC-P
(1)
All Applications
Filter By Species
Human
(12)
Mouse
(2)
N/A
(1)
All Species
Showing Publications 1 - 10 of 16. Show All 16 Publications.
Publications using 707-C3 Applications Species
Grin, PM;Baid, K;de Jesus, HCR;Kozarac, N;Bell, PA;Jiang, SZ;Kappelhoff, R;Butler, GS;Leborgne, NGF;Pan, C;Pablos, I;Machado, Y;Vederas, JC;Kim, H;Benarafa, C;Banerjee, A;Overall, CM; SARS-CoV-2 3CLpro (main protease) regulates caspase activation of gasdermin-D/E pores leading to secretion and extracellular activity of 3CLpro Cell reports 2024-12-12 [PMID: 39673710] (Bioassay, Human) Bioassay Human
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 Genuine selective caspase-2 inhibition with new irreversible small peptidomimetics Cell Death & Disease, 2022-11-15;13(11):959. 2022-11-15 [PMID: 36379916] (Bioassay, N/A) Bioassay N/A
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 The metabolite &amp;alpha-KG induces GSDMC-dependent pyroptosis through death receptor 6-activated caspase-8 Cell Research, 2021-05-19;0(0):. 2021-05-19 [PMID: 34012073] (Bioassay, Human) Bioassay Human
H Shen, H Zhu, D Panja, Q Gu, Z Li Autophagy controls the induction and developmental decline of NMDAR-LTD through endocytic recycling Nat Commun, 2020-06-12;11(1):2979. 2020-06-12 [PMID: 32532981] (Bioassay, Mouse) Bioassay Mouse
D Huo, J Zhu, G Chen, Q Chen, C Zhang, X Luo, W Jiang, X Jiang, Z Gu, Y Hu Eradication of unresectable liver metastasis through induction of tumour specific energy depletion Nat Commun, 2019-07-11;10(1):3051. 2019-07-11 [PMID: 31296864] (Bioassay, Human) Bioassay Human
YJ Lee, MS Shu, JY Kim, YH Kim, KH Sim, WJ Sung, JR Eun Cilostazol protects hepatocytes against alcohol-induced apoptosis via activation of AMPK pathway PLoS ONE, 2019-01-29;14(1):e0211415. 2019-01-29 [PMID: 30695051] (Bioassay) Bioassay
X Qin, M Lyu, Y Si, J Yang, Z Wu, J Li Alkyne-based surface-enhanced Raman scattering nanoprobe for ratiometric imaging analysis of caspase 3 in live cells and tissues Anal. Chim. Acta, 2018-09-08;1043(0):115-122. 2018-09-08 [PMID: 30392659] (Bioassay, Human) Bioassay Human
B Zhou, JY Zhang, XS Liu, HZ Chen, YL Ai, K Cheng, RY Sun, D Zhou, J Han, Q Wu Tom20 senses iron-activated ROS signaling to promote melanoma cell pyroptosis Cell Res., 2018-10-04;0(0):. 2018-10-04 [PMID: 30287942] (Bioassay, Human) Bioassay Human
F Wang, L Wang, J Wu, I Sokirniy, P Nguyen, T Bregnard, J Weinstock, M Mattern, I Bezsonova, WW Hancock, S Kumar Active site-targeted covalent irreversible inhibitors of USP7 impair the functions of Foxp3+ T-regulatory cells by promoting ubiquitination of Tip60 PLoS ONE, 2017-12-13;12(12):e0189744. 2017-12-13 [PMID: 29236775] (Bioassay, Human) Bioassay Human
MK Shim, HY Yoon, S Lee, MK Jo, J Park, JH Kim, SY Jeong, IC Kwon, K Kim Caspase-3/-7-Specific Metabolic Precursor for Bioorthogonal Tracking of Tumor Apoptosis Sci Rep, 2017-11-30;7(1):16635. 2017-11-30 [PMID: 29192289] (Enzyme Assay, Human) Enzyme Assay Human
Show All 16 Publications.

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Blogs on Caspase-3. Showing 1-10 of 11 blog posts - Show all blog posts.

Pyroptosis: Mechanisms mediating cell death and pro-inflammatory cytokine release
By Victoria OsinskiPyroptosis is an inflammatory form of programmed cell death characterized by the release of pro-inflammatory cytokines IL-1 beta and IL-18.1,10 It is a process distinct from apoptosis and necrosis (T...  Read full blog post.

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By Rachel M.A. Linger, Ph.D.What is survivin?Survivin is a small (16.5 kDa) protein normally found in human fetal tissue. In contrast, survivin is typically undetectable in most normal adult tissues. Expression of ...  Read full blog post.

Cleaved Caspase-3: A Marker of Programmed Cell Death
What are Caspases?Caspases, or cysteine-dependent aspartate specific proteases, are a family of enzymes crucial for initiating and executing apoptosis within a cell, an important biological event especially during organ development (1). Environme...  Read full blog post.

Caspase-3, The Executioner of Apoptosis
The Role of Caspase-3 in ApoptosisCaspase-3 enzyme is a member of the family of endoproteases which regulate inflammation and apoptosis signaling networks. Caspase-3 is known as an executioner caspase in apoptosis because of its role in coordinat...  Read full blog post.

Caspase 7 - A key effector of the apoptotic pathway
Caspase-7 is an effector caspase with important roles in mediating cell death signaling. As an effector caspase, caspase-7 is cleaved and activated by initiator caspases such as caspase-1 (1). Like other caspase family proteins, caspase-7 contains a...  Read full blog post.

Caspase 11 - A proinflammatory caspase that induces the innate immune response
While known for their role in programmed cell death, caspases are also essential for mediating inflammatory responses and innate immunity. Binding of microbial molecules by pattern recognition receptors triggers the formation of the multiprotein in...  Read full blog post.

LC3/LC3B - measuring autophagosome formation and autophagic flux
Microtubule-associated protein-1 light chain 3 (LC3/LC3B) is a ubiquitin-like protein involved in the formation of the autophagosome. It is homologous to the yeast Atg8 protein. Autophagosomes are important for the degradation and recycling of intr...  Read full blog post.

D4-GDI (GDP dissociation inhibitor, RhoGD12)
The D4-GDI protein is a negative regulator of the Ras-related Rho family of small molecule "molecular switch" GTPases. The Rho GTPases modify cell structure and architecture via rapid changes to the actin cytoskeleton and cell membrane. M...  Read full blog post.

Showing 1-10 of 11 blog posts - Show all blog posts.

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Bioinformatics

Gene Symbol CASP3
Uniprot