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Caspase-3/CASP3 Protein, Human

6 Cited Publications
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Based on 6 publication(s) in Google Scholar

Caspase-3/CASP3 protein is a thiol protease critical in apoptosis and is activated by initiating caspases (CASP8, CASP9 and/or CASP10). It catalyzes the cleavage of multiple proteins in sympathetic neurons, including PARP1, SREBP, caspase-6, -7 and -9, huntingtin, and RET. Caspase-3/CASP3 Protein, Human is the recombinant human-derived Caspase-3/CASP3 protein, expressed by E. coli, with tag free.

For research use only. We do not sell to patients.
  • Species: Human
  • Source: E. coli
  • Storage:
    Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.
  • Experimental Validation Results for Caspase-3/CASP3 Protein, Human
  • Experimental Validation Results for Caspase-3/CASP3 Protein, Human
  • Experimental Validation Results for Caspase-3/CASP3 Protein, Human
  • Caspase-3/CASP3 Protein, Human Product Image
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To place orders, for customer services and technical support, please contact: MedChemExpress USA

Tel: 609-228-6898 E-mail: [email protected] [email protected]

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To place orders, for customer services and technical support, please contact:

609-228-6898
[email protected] [email protected]
Find Your Local Distributor Your local distributor: Hycultec GmbHHycultec
  • Biological Activity
  • Technical Parameters
  • Product Properties
  • Documentation
  • Help & FAQs
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Biological Activity

Description

Caspase-3/CASP3 protein is a thiol protease critical in apoptosis and is activated by initiating caspases (CASP8, CASP9 and/or CASP10). It catalyzes the cleavage of multiple proteins in sympathetic neurons, including PARP1, SREBP, caspase-6, -7 and -9, huntingtin, and RET. Caspase-3/CASP3 Protein, Human is the recombinant human-derived Caspase-3/CASP3 protein, expressed by E. coli, with tag free.

Background

Caspase-3/CASP3 Protein functions as a pivotal thiol protease in the execution phase of apoptosis, serving as a major effector caspase. Upon cleavage and activation by initiator caspases (CASP8, CASP9, and/or CASP10), it orchestrates apoptosis by catalyzing the cleavage of numerous proteins. In the early stages of apoptosis, it proteolytically cleaves poly(ADP-ribose) polymerase PARP1, targeting the '216-Asp-|-Gly-217' bond. CASP3 also activates sterol regulatory element-binding proteins (SREBPs), cleaves and activates caspase-6, -7, and -9, participates in the cleavage of huntingtin, and induces cell adhesion in sympathetic neurons through RET cleavage. Additionally, CASP3 cleaves and inhibits serine/threonine-protein kinase AKT1 in response to oxidative stress. It acts as an inhibitor of type I interferon production during virus-induced apoptosis by cleaving antiviral proteins CGAS, IRF3, and MAVS, thus preventing cytokine overproduction. CASP3 is also involved in pyroptosis, mediating the cleavage and activation of gasdermin-E (GSDME). Furthermore, it cleaves XRCC4 and phospholipid scramblase proteins XKR4, XKR8, and XKR9, promoting phosphatidylserine exposure on the apoptotic cell surface.

Verified Bioactivity

Measured by its ability to cleave the fluorogenic peptide substrate Ac-DEVD-AFC. The specific activity is ≥26 U/mg, as measured under the described conditions.
1U: The amount of enzyme required to consume l μmol of substrate per minute at 25°C.

Assay Procedure

Materials
Assay buffer: 25 mM HEPES, 0.1% (w/v) CHAPS, 10 mM dithiothreitol (DTT) , pH 7.5
Test protein: Caspase-3/CASP3 Protein, Human (HY-P701341)
Substrate: Ac-DEVD-AFC (HY-P1005), stored as a 10 mM solution diluted in DMSO.
Standard：7-amino, 4-(trifluoromethyl) coumarin (HY-D0981)

Procedure
1. Dilute 7-amino,4-(trifluoromethyl) coumarin in assay buffer to concentrations of 0, 1.5625, 3.125, 6.25, 12.5, 25, 50, and 100 μmol/L. Add 100 μL of each dilution to a black microplate well. Measure fluorescence at excitation and emission wavelengths of 400 nm and 505 nm, respectively, in kinetic mode for 5 minutes. Plot the measured RFU values on the y-axis and the standard concentrations on the x-axis to generate a standard curve and obtain the curve equation.
2. Dilute Human Caspase 3 to 0.4 μg/mL in assay buffer.
3. Dilute the substrate to 100 μM in assay buffer.
4. Experimental group: Add 50 μL of 0.4 μg/mL Human Caspase 3 to the plate, then add 50 μL of 100 μM substrate to initiate the reaction.
Control group: Add 50 μL of assay buffer and 50 μL of 100 μM substrate.
5. Read in kinetic mode for 5 minutes at excitation and emission wavelengths of 400 nm and 505 nm, respectively.
6. 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

Technical Parameters

  • Species Human
  • Source E. coli
  • Tag Tag Free
  • Accession

    P42574 (S29-D175&S176-H277)

  • Gene ID
    836
  • Molecular Construction
    • N-term
    • CASP3 (S29-D175&S176-H277)
      Accession # P42574
    • C-term
  • Protein Length

    Full Length of Mature Protein

  • Synonyms

    CASP3; Protein Yama; Caspase 3; Caspase-3; Apopain; CASP-3; CPP32; CPP-32; CPP32B; SCA-1; Caspase 3, Apoptosis-Related Cysteine Peptidase; Yama; Caspase 3, Apoptosis-Related Cysteine Protease; PARP Cleavage Protease; SREBP Cleavage Activity 1; Procaspase3

  • Predicted Molecular Mass

    12.6 kDa & 16.6 kDa

  • Molecular Weight

    Approximately 12 kDa & 16 kDa, based on SDS-PAGE under reducing conditions.

  • Purity

    ≥ 90%, as determined by reducing SDS-PAGE.

Product Properties

Appearance

Solution

Formulation

Supplied as a 0.22 μm filtered solution of 50 mM Tris-HCl, pH 7.5, 200 mM NaCl, 20% glycerol, 1 mM DTT.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

Please use rapid thawing with running water to thaw the protein.

Storage & Stability

Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping

Shipping with dry ice.

Calculators

Reconstitution Calculator

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration

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Dilution Calculator

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

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The Specific Activity Calculator Equation
  • Specific Activity (Unit/mg)
  • Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) Specific Activity (Unit/mg)
Unit/mg
= 106 ÷
Biological Activity (ED50) Biological Activity (ED50)
106 ÷
ng/mL
MOQ
Minimum order quantity
100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    Caspase-3/CASP3 Protein, Human purchased from MedChemExpress. Usage Cited in: Sci Adv. 2025 Aug;11(31):eadw5133. [Abstract]

    Customer Validation
     Cumulative drug release curves of HCeOx-­D@PM in the presence of PBS (pH 7.4), 1 mM H2O2, and Caspase-3/CASP3 Protein, Human (Casp3; 0.1 and 1 μg/mL) at specific time points.

  • 1/16

    Caspase-3/CASP3 Protein, Human purchased from MedChemExpress. Usage Cited in: Sci Adv. 2025 Aug;11(31):eadw5133. [Abstract]

    Customer Validation
     Cumulative drug release and corresponding fitting of HCeOx-­D@PM in the presence of different Caspase-3/CASP3 Protein, Human (Casp3; 0, 0.5, 1, 1.5, 2 μg/mL) concentrations over 24 hours.

  • 1/16

    Caspase-3/CASP3 Protein, Human purchased from MedChemExpress. Usage Cited in: RSC Adv. 2024 Nov 18;14(49):36754-36762. [Abstract]

    Customer Validation
     SDS-PAGE analysis of papain, Caspase-3/CASP3 Protein, Human (caspase-3), and thioredoxin (TXN) proteins after treatment with compound 5.

  • Caspase-3/CASP3 Protein, Human Product Image
    1/16
  • Experimental Validation Results for Caspase-3/CASP3 Protein, Human
  • Experimental Validation Results for Caspase-3/CASP3 Protein, Human
  • Experimental Validation Results for Caspase-3/CASP3 Protein, Human
  • Caspase-3/CASP3 Protein, Human Product Image

  • Caspase-3/CASP3 Protein, Human purchased from MedChemExpress. Usage Cited in: Sci Adv. 2025 Aug;11(31):eadw5133. [Abstract]

    Cumulative drug release curves of HCeOx-­D@PM in the presence of PBS (pH 7.4), 1 mM H2O2, and Caspase-3/CASP3 Protein, Human (Casp3; 0.1 and 1 μg/mL) at specific time points.

  • Caspase-3/CASP3 Protein, Human purchased from MedChemExpress. Usage Cited in: Sci Adv. 2025 Aug;11(31):eadw5133. [Abstract]

    Cumulative drug release and corresponding fitting of HCeOx-­D@PM in the presence of different Caspase-3/CASP3 Protein, Human (Casp3; 0, 0.5, 1, 1.5, 2 μg/mL) concentrations over 24 hours.

  • Caspase-3/CASP3 Protein, Human purchased from MedChemExpress. Usage Cited in: RSC Adv. 2024 Nov 18;14(49):36754-36762. [Abstract]

    SDS-PAGE analysis of papain, Caspase-3/CASP3 Protein, Human (caspase-3), and thioredoxin (TXN) proteins after treatment with compound 5.