Recombinant Mouse Cathepsin B Protein, CF

R&D Systems | Catalog # 965-CY

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Cathepsin B Protein (965-CY)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

P10605

Structure / Form

Pro and mature forms

Applications

Enzyme Activity
View all Cathepsin B Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Cathepsin B protein
His18-Phe339, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

His18

Predicted Molecular Mass

37 kDa

SDS-PAGE

43 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate Z-LR-AMC (Catalog # ES008).
The specific activity is >2,000 pmol/min/µg, as measured under the described conditions. 

Formulation, Preparation, and Storage

965-CY
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with polar packs. 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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Cathepsin B

Cathepsin B is the first described member of the family of lysosomal cysteine proteases (1). Cathepsin B possesses both endopeptidase and exopeptidase activities, in the latter case acting as a peptidyl-dipeptidase. It is known to process a number of proteins, including pro and active caspases, prorenin and secretory leucoprotease inhibitor (SLPI) (2-4). Therefore, Cathepsin B may play a role in activation and inactivation of caspases, activation of renin and inactivation of SLPI, the key steps in apoptosis, angiotensin production, and progression of emphysema, respectively. Because of its increased levels and redistribution in human and animal tumors, Cathepsin B may also have a role in invasion and metastasis (5). In addition to the lysosome, Cathepsin B can be secreted or associated with plasma membrane, cytoplasm, and nucleus. It is synthesized as a preproenzyme. Following removal of the signal peptide, the inactive proenzyme undergoes further modifications including removal of the pro region to result in the active enzyme (5).

References

  1. Mort, J.S. (2004) in Handbook of Proteolytic Enzymes (Barrett, A.J. et al. eds.) p. 1079, Academic Press, San Diego.
  2. Vancompernolle, K. et al. (1998) FEBS Lett. 438:150.
  3. Jutras, I. and T.L. Reudelhuber (1998) FEBS Lett. 443:48.
  4. Taggart, C.C. et al. (2001) J. Biol. Chem. 276:33345.
  5. Berquin, I.M. and B.F. Sloane (1996) Adv. Exp. Med. Biol. 389:281.

Alternate Names

CTSB

Entrez Gene IDs

1508 (Human); 13030 (Mouse)

Gene Symbol

CTSB

UniProt

P10605

Additional Cathepsin B Products

Product Documents for Recombinant Mouse Cathepsin B Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse Cathepsin B Protein, CF

For research use only

Citations for Recombinant Mouse Cathepsin B Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Cathepsin B Protein, CF (965-CY):

Materials
  • Activation Buffer: 25 mM MES, 5 mM DTT, pH 5.0
  • Assay Buffer: 25 mM MES, pH 5.0
  • Recombinant Mouse Cathepsin B (rmCathepsin B) (Catalog # 965-CY)
  • Substrate: Z-Leu-Arg-AMC (Catalog # ES008), 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 rmCathepsin B to 10 µg/mL in Activation Buffer.
  2. Incubate at room temperature for 15 minutes (activation step).
  3. Dilute activated rmCathepsin B to 0.2 ng/µL in Assay Buffer.
  4. Dilute Substrate 20 µM in Assay Buffer.
  5. In a plate load 50 µL of 0.2 ng/µL rmCathepsin B to wells, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank of 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
  6. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. 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-Methyl Coumarin (AMC) (Sigma, Catalog # A-9891).

Per Well:

  • rmCathepsin B: 0.01 µg
  • Substrate: 10 µM

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