Recombinant Mouse Cathepsin B Protein, CF

Catalog # Availability Size / Price Qty
965-CY-010
Product Details
Citations (4)
FAQs
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Recombinant Mouse Cathepsin B Protein, CF Summary

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.
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. 
Source
Mouse myeloma cell line, NS0-derived mouse Cathepsin B protein
His18-Phe339, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
His18
Structure / Form
Pro and mature forms
Predicted Molecular Mass
37 kDa
SDS-PAGE
43 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

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.

Assay Procedure

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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.
Entrez Gene IDs
1508 (Human); 13030 (Mouse)
Alternate Names
APP secretase; APPS; Cathepsin B; Cathepsin B1; CPSBamyloid precursor protein secretase; CTSB; cysteine protease; EC 3.4.22; EC 3.4.22.1

Citations for Recombinant Mouse Cathepsin B Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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  1. Modulation of Receptor Protein Tyrosine Phosphatase Sigma Increases Chondroitin Sulfate Proteoglycan Degradation through Cathepsin B Secretion to Enhance Axon Outgrowth
    Authors: AP Tran, S Sundar, M Yu, BT Lang, J Silver
    J. Neurosci., 2018;38(23):5399-5414.
    Species: Rat
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Cathepsins limit macrophage necroptosis through cleavage of Rip1 kinase.
    Authors: McComb S, Shutinoski B, Thurston S, Cessford E, Kumar K, Sad S
    J Immunol, 2014;192(12):5671-8.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  3. Chemotherapy-triggered cathepsin B release in myeloid-derived suppressor cells activates the Nlrp3 inflammasome and promotes tumor growth.
    Authors: Bruchard M, Mignot G, Derangere V, Chalmin F, Chevriaux A, Vegran F, Boireau W, Simon B, Ryffel B, Connat J, Kanellopoulos J, Martin F, Rebe C, Apetoh L, Ghiringhelli F
    Nat Med, 2012;19(1):57-64.
    Species: Mouse
    Sample Types: N/A
    Applications: Binding Assay
  4. VEGF-A induces angiogenesis by perturbing the cathepsin-cysteine protease inhibitor balance in venules, causing basement membrane degradation and mother vessel formation.
    Authors: Chang SH, Kanasaki K, Gocheva V, Blum G, Harper J, Moses MA, Shih SC, Nagy JA, Joyce J, Bogyo M, Kalluri R, Dvorak HF
    Cancer Res., 2009;69(10):4537-44.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo

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Fluorogenic Peptide Substrates

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