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Recombinant Human Cathepsin B Protein, CF

R&D Systems | Catalog # 953-CY

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Cathepsin B Protein (953-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

P07858

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 human Cathepsin B protein
Arg18-Ile339 (pro) & Phe74-Ile339 (mature), both 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

Arg18 & Phe74

Predicted Molecular Mass

37 kDa (Pro) & 29 kDa (Mature)

SDS-PAGE

43 kDa and 36 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate Z-LR-AMC (Catalog # ES008).
The specific activity is >2,500 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

953-CY
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with dry ice or equivalent. 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 of the enzyme in human and animal tumors, Cathepsin B may also have role in invasion and metastasis (5).

In addition to 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 (1).

References

  1. Mort, J.S. (2004) in Handbook of Proteolytic Enzymes. Barrett, A.J. et al. (eds): Academic Press, San Diego, p. 1079.
  2. Vancompernolle, K. et al. (1998) FEBS Lett. 438:150.
  3. Jutras, I. and T.L. Reudelhuber (1999) FEBS Lett. 443:48.
  4. Taggart, C.C. et al. (2001) J. Biol. Chem. 276:33345.
  5. Bergquin, 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

P07858

Additional Cathepsin B Products

Product Documents for Recombinant Human Cathepsin B Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human Cathepsin B Protein, CF

For research use only

Citations for Recombinant Human Cathepsin B Protein, CF

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Protocols

View specific protocols for Recombinant Human Cathepsin B Protein, CF (953-CY):

Materials
  • Activation Buffer: 25 mM MES, 5 mM DTT, pH 5.0
  • Assay Buffer: 25 mM MES,0.02% Brij-35, pH 5.0
  • Recombinant Human Cathepsin B (rhCathepsin B) (Catalog # 953-CY)
  • Fluorogenic Peptide Substrate VII: Z-Leu-Arg-AMC (Catalog # ES008)
  • Black 96-well Plate
  • Plate Reader with Fluorescence Read Capability
  1. Dilute rhCathepsin B to 10 µg/mL in Activation Buffer.
  2. Incubate at room temperature for 15 minutes.
  3. Dilute rhCathepsin B to 0.1 µg/mL in Assay Buffer.
  4. Dilute substrate to 80 µM in Assay Buffer.
  5. Load 50 µL of the 0.1 µg/mL rhCathepsin B in a black well plate, and start the reaction by adding 50 µL of 80 µM Substrate.  Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 80 µM Substrate.
  6. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
    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).

Per Well:

  • rhCathepsin B: 0.005 µg
  • Substrate: 40 µM

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