Cathepsin B Protein, Rat (HEK293, His)
Based on 1 Customer Validation
Cathepsin B is a lysosomal cysteine protease that plays a role in intracellular protein catabolism. Cathepsin B mediates JNK signaling pathway to regulate the migration of glioma initiation cells. Cathepsin B is involved in the pathology of chronic inflammatory diseases of the airway and joints, as well as cancer and pancreatitis. Cathepsin B Protein, Rat (HEK293, His) is the recombinant rat-derived Cathepsin B protein, expressed by HEK293 , with C-His labeled tag.
- Species: Rat
- Source: HEK293
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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.
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Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Technical Parameters
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Product Properties
- Documentation
- Help & FAQs
Biological Activity
Description
Cathepsin B is a lysosomal cysteine protease that plays a role in intracellular protein catabolism. Cathepsin B mediates JNK signaling pathway to regulate the migration of glioma initiation cells. Cathepsin B is involved in the pathology of chronic inflammatory diseases of the airway and joints, as well as cancer and pancreatitis. Cathepsin B Protein, Rat (HEK293, His) is the recombinant rat-derived Cathepsin B protein, expressed by HEK293 , with C-His labeled tag.
Background
Cathepsin B is a lysosomal cysteine protease that plays a role in intracellular protein catabolism and is involved in other physiological processes such as antigen processing in immune response, hormone activation, and bone turnover. Cathepsin B mediates JNK signaling pathway to regulate the migration of glioma initiation cells. Cathepsin B can enhance the activity of other proteases, including matrix metalloproteinases, urokinase (serine protease urokinase plasminogen activator), and Cathepsin D. Cathepsin B is involved in the pathology of chronic inflammatory diseases of the airway and joints, as well as cancer and pancreatitis[1][2][3].
Verified Bioactivity
Measured by its ability to cleave the fluorogenic peptide substrate Z-LR-AMC . The specific activity is >2,500 pmol/min/μg. (Activation description: The proenzyme needs to be activated in acid reducing buffer for an activated form.)
MCE Validation Data
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Purity - SDS-PAGE
Purity - SDS-PAGE
Assay Procedure
Materials
Activation buffer: 25 mM MES, 5 mM DTT, pH 5.0
Assay buffer: 25 mM MES, pH 5.0
Cathepsin B Protein, Rat (HEK293, His) (HY-P74345)
Substrate: Z-Leu-Arg-AMC (HY-142021)
Standard: 7-Amino-4-methylcoumarin (AMC, HY-D0027)
Procedure
1. Dilute AMC to 0, 0.78125, 1.5625, 3.125, 6.25, 12.5, 25, 50, and 100 μmol/L respectively with assay buffer.
2. Add 100 μL of each dilution to black microplate wells.
3. Set excitation and emission wavelengths to 380 nm and 460 nm (top read), and read in kinetic mode for 5 minutes.
4. Plot a standard curve with the measured RFU as the abscissa and the amount of standard substance as the ordinate, and obtain the standard curve equation.
5. Dilute the test protein to 10 μg/mL with activation buffer; dilute Rat Cathepsin B to 100 μg/mL with assay buffer.
6. Incubate at room temperature for 15 minutes.
7. Dilute Rat Cathepsin B to 0.2 μg/mL with assay buffer.
8. Dilute the substrate to 20 μM with assay buffer.
9. Add 50 μL of the test protein at each concentration to the assay wells, and initiate the reaction by adding 50 μL of 20 μM substrate. For blank wells, add 50 μL of assay buffer and 50 μL of 20 μM substrate without protein.
10. Set excitation and emission wavelengths to 380 nm and 460 nm (top read), and read in kinetic mode for 5 minutes.
11. Calculate the 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
Per Well:
Rat Cathepsin B: 0.01 μg
Substrate: 10 μM
Technical Parameters
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Species Rat
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Source HEK293
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Tag C-6*His
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Accession
Q6IN22 (H18-F339)
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Molecular Construction
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N-term
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Cathepsin B (H18-F339)
Accession # Q6IN22 -
His
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C-term
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Protein Length
Full Length of Mature Protein
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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
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AA Sequence
HDKPSFHPLSDDMINYINKQNTTWQAGRNFYNVDISYLKKLCGTVLGGPKLPERVGFSEDINLPESFDAREQWSNCPTIAQIRDQGSCGSCWAFGAVEAMSDRICIHTNGRVNVEVSAEDLLTCCGIQCGDGCNGGYPSGAWNFWTRKGLVSGGVYNSHIGCLPYTIPPCEHHVNGSRPPCTGEGDTPKCNKMCEAGYSTSYKEDKHYGYTSYSVSDSEKEIMAEIYKNGPVEGAFTVFSDFLTYKSGVYKHEAGDVMGGHAIRILGWGIENGVPYWLVANSWNVDWGDNGFFKILRGENHCGIESEIVAGIPRTQQYWGRF
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Molecular Weight
Approximately 35-43 kDa due to the glycosylation.
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Purity
≥ 95%, as determined by reducing SDS-PAGE.
Product Properties
Lyophilized powder.
1.Lyophilized from a 0.22 μm filtered solution of 25 mM Tris, 150 mM NaCl, pH 7.5, 5% trehalose, 5% mannitol, 0.01% Tween 80.
2.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4.
Please refer to the lot-specific COA for specific buffer information.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.
<1 EU/μg, determined by LAL method.
It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O.
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.
Room temperature in continental US; may vary elsewhere.
Documentation
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Data Sheet (238 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Handling Instructions (2659 KB)
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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