Copy Link Print Information

Cathepsin B Protein, Human (HEK293, His)

2 Cited Publications
Customer Review

Based on 2 publication(s) in Google Scholar

Cathepsin B Protein, Human (HEK293, His) functions in intracellular protein catabolism and in certain situations may also be involved in other physiological processes, such as processing of antigens in the immune response, hormone activation and bone turnover.

For research use only. We do not sell to patients.
  • Species: Human
  • Source: HEK293
  • Storage:
    Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.
  • Experimental Validation Results for Cathepsin B Protein, Human (HEK293, His)
  • Experimental Validation Results for Cathepsin B Protein, Human (HEK293, His)
  • Cathepsin B Protein, Human (HEK293, His) Product Image
Size
Price
Stock
Quantity
Amount: USD 0.00
Please select quantity
or
or

* Please select Quantity before adding items.

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

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

Service
Data Download

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
Top Publications Citing

Use of MCE Products

Biological Activity

Description

Cathepsin B Protein, Human (HEK293, His) functions in intracellular protein catabolism and in certain situations may also be involved in other physiological processes, such as processing of antigens in the immune response, hormone activation and bone turnover[1].

Background

Cathepsin B is a lysosomal cysteine protease of the papain family. It functions in intracellular protein catabolism and in certain situations may also be involved in other physiological processes, such as processing of antigens in the immune response, hormone activation and bone turnover. There is also evidence that cathepsin B is implicated in the pathology of chronic inflammatory diseases of airways and joints, and in cancer and pancreatitis[1].

Verified Bioactivity

Measured by its ability to cleave the fluorogenic peptide substrate Z-LR-AMC. Read at excitation and emission wavelengths of 380 nm and 460 nm. The specific activity is 6500-20000 pmol/min/µg, as measured under the described conditions. (Activation description: The proenzyme needs to be activated in acid reducing buffer for an activated form.)

MCE Validation Data

  • Purity - SDS-PAGE

    Purity - SDS-PAGE

    ≥ 95%, as determined by reducing SDS-PAGE.

  • Bioactivity - Enzymatic Activity Assay

    Bioactivity - Enzymatic Activity Assay

    Measured by its ability to cleave the fluorogenic peptide substrate Z-LR-AMC. Read at excitation and emission wavelengths of 380 nm and 460 nm. The specific activity is 8552.8807 pmol/min/µg, as measured under the described conditions.

Assay Procedure

Materials
Activation buffer: 25 mM MES, 5 mM DTT, pH 5.0
Assay buffer: 25 mM MES, 0.02% Brij-35, pH 5.0
Test protein: Cathepsin B Protein, Human (HEK293, C-His) (HY-P7993A)
Substrate: Z-Leu-Arg-AMC (HY-142021)
Standard: 7-amino, 4-Methyl Coumarin (AMC, HY-D0027)

Procedure
1. Standard Curve: Dilute AMC in assay buffer to concentrations of 0, 0.4, 0.8, 1.2, 1.6, 2, 6.25, 12.5, 25, 50, and 100 μmol/L, respectively. Add 100 μL of each dilution into black microplate wells. Set the excitation and emission wavelengths to 380 nm and 460 nm, respectively. Plot the standard curve with the measured RFU (Relative Fluorescence Units) on the x-axis and the amount of standard substance on the y-axis to obtain the standard curve equation.
2. Dilute the protein sample to 10 μg/mL in activation buffer.
3. Incubate at room temperature for 15 minutes.
4. Dilute Human Cathepsin B to 0.2 μg/mL in assay buffer.
5. Dilute the substrate to 20 μM in assay buffer.
6. Add 50 μL of each protein concentration into the assay wells and add 50 μL of 20 μM substrate to initiate the reaction. For the blank wells, add 50 μL of assay buffer and 50 μL of 20 μM substrate without protein.
7. Set the excitation and emission wavelengths to 380 nm and 460 nm, respectively. Read the fluorescence in kinetic mode for 5 minutes.
8. Calculate Specific Activity:

     Specific Activity (pmol/min/μg) =

Adjusted Vmax* (RFU/min) x Conversion Factor (pmol/RFU) **
amount of enzyme (μg)
*Adjusted for Control
**Derived using calibration standard AMC.

Technical Parameters

  • Species Human
  • Source HEK293
  • Tag C-6*His
  • Accession

    P07858 (R18-I339)

  • Gene ID
    1508
  • Molecular Construction
    • N-term
    • Cathepsin B (R18-I339)
      Accession # P07858
    • 6*His
    • C-term
  • Protein Length

    Full Length of Mature Protein

  • Synonyms

    CTSB; Keratolytic Winter Erythema (Oudtshoorn Skin Disease); Cathepsin B; Epididymis Secretory Sperm Binding Protein; APP Secretase; Amyloid Precursor Protein Secretase; Cathepsin B1; Cysteine Protease; APPS; RECEUP; CPSB; KWE

  • AA Sequence

    RSRPSFHPLSDELVNYVNKRNTTWQAGHNFYNVDMSYLKRLCGTFLGGPKPPQRVMFTEDLKLPASFDAREQWPQCPTIKEIRDQGSCGSCWAFGAVEAISDRICIHTNAHVSVEVSAEDLLTCCGSMCGDGCNGGYPAEAWNFWTRKGLVSGGLYESHVGCRPYSIPPCEHHVNGSRPPCTGEGDTPKCSKICEPGYSPTYKQDKHYGYNSYSVSNSEKDIMAEIYKNGPVEGAFSVYSDFLLYKSGVYQHVTGEMMGGHAIRILGWGVENGTPYWLVANSWNTDWGDNGFFKILRGQDHCGIESEVVAGIPRTDQYWEKI

  • Predicted Molecular Mass

    36.7 kDa

  • Molecular Weight

    Approximately 37-45 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.

  • Glycosylation

    Yes

  • Purity

    ≥ 95%, as determined by reducing SDS-PAGE.

Product Properties

Appearance

Lyophilized powder

Formulation

1.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4.
2.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 8% trehalose.
Please refer to the lot-specific COA for specific buffer information.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Storage & Stability

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Shipping

Room temperature in continental US; may vary elsewhere.

Calculators

Reconstitution Calculator

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

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

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

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

Get Quote In-stock

Other size
Get Quote
Please select quantity
Amount: USD 0.00

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

    MedChemExpress Validation
     ≥ 95%, as determined by reducing SDS-PAGE.

  • 1/16

    MedChemExpress Validation
     Measured by its ability to cleave the fluorogenic peptide substrate Z-LR-AMC. Read at excitation and emission wavelengths of 380 nm and 460 nm. The specific activity is 8552.8807 pmol/min/µg, as measured under the described conditions.

  • Cathepsin B Protein, Human (HEK293, His) Product Image
    1/16
  • Experimental Validation Results for Cathepsin B Protein, Human (HEK293, His)
  • Experimental Validation Results for Cathepsin B Protein, Human (HEK293, His)
  • Cathepsin B Protein, Human (HEK293, His) Product Image