Recombinant Human Cathepsin B Protein, CF

Catalog # Availability Size / Price Qty
953-CY-010
Product Details
Citations (11)
FAQs
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Recombinant Human 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 >2,500 pmol/min/µg, as measured under the described conditions.
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
Accession #
N-terminal Sequence
Analysis
Arg18 & Phe74
Structure / Form
Pro and Mature forms
Predicted Molecular Mass
37 kDa (Pro) & 29 kDa (Mature)
SDS-PAGE
43 kDa and 36 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.

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.

Assay Procedure

Materials
  • Activation Buffer: 25 mM MES, 5 mM DTT, pH 5.0
  • Assay Buffer: 25 mM MES, pH 5.0
  • Recombinant Human Cathepsin B (rhCathepsin B) (Catalog # 953-CY)
  • Fluorogenic Peptide Substrate VII: Z-Leu-Arg-AMC (Catalog # ES008)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhCathepsin B to 10 µg/mL in Activation Buffer.
  2. Incubate at room temperature for 15 minutes.
  3. Dilute rhCathepsin B to 0.2 ng/µL in Assay Buffer.
  4. Dilute substrate to 20 µM in Assay Buffer.
  5. Load 50 µL of the 0.2 ng/µL rhCathepsin B in a black well plate, and start the reaction by adding 50 µL of 20 µM Substrate.  Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate without any rhCathepsin B.
  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) (Sigma, Catalog # A-9891).

Per Well:
  • rhCathepsin B: 0.01 µg
  • Substrate: 10 µM
Reconstitution Calculator

Reconstitution Calculator

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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 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.
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 Human 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.

11 Citations: Showing 1 - 10
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  1. Human Cysteine Cathepsins Degrade Immunoglobulin G In Vitro in a Predictable Manner
    Authors: RA Høglund, SB Torsetnes, A Lossius, B Bogen, EJ Homan, R Bremel, T Holmøy
    Int J Mol Sci, 2019;20(19):.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  2. Eradication of unresectable liver metastasis through induction of tumour specific energy depletion
    Authors: D Huo, J Zhu, G Chen, Q Chen, C Zhang, X Luo, W Jiang, X Jiang, Z Gu, Y Hu
    Nat Commun, 2019;10(1):3051.
    Species: Human
    Sample Types: Nanoparticles
    Applications: Bioassay
  3. The lysosomal protein cathepsin L is a progranulin protease
    Authors: CW Lee, JN Stankowski, J Chew, CN Cook, YW Lam, S Almeida, Y Carlomagno, KF Lau, M Prudencio, FB Gao, M Bogyo, DW Dickson, L Petrucelli
    Mol Neurodegener, 2017;12(1):55.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  4. Granzyme B PET Imaging as a Predictive Biomarker of Immunotherapy Response
    Authors: BM Larimer, E Wehrenberg, F Dubois, A Mehta, T Kalomeris, K Flaherty, G Boland, U Mahmood
    Cancer Res., 2017;77(9):2318-2327.
    Species: N/A
    Sample Types: Protein
    Applications: Bioassay
  5. Cathepsin K-mediated Notch1 activation contributes to neovascularization in response to hypoxia.
    Authors: Jiang H, Cheng X, Shi G, Hu L, Inoue A, Yamamura Y, Wu H, Takeshita K, Li X, Huang Z, Song H, Asai M, Hao C, Unno K, Koike T, Oshida Y, Okumura K, Murohara T, Kuzuya M
    Nat Commun, 2014;5(0):3838.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Bioassay
  6. Endosomal acidification and cathepsin L activity is required for calicivirus replication.
    Authors: Shivanna V, Kim Y, Chang K
    Virology, 2014;464(0):287-95.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  7. 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: Human
    Sample Types: N/A
    Applications: Surface Plasmon Resonance
  8. Fine-tuning nucleophosmin in macrophage differentiation and activation.
    Authors: Guery L, Benikhlef N, Gautier T, Paul C, Jego G, Dufour E, Jacquel A, Cally R, Manoury B, Vanden Berghe T, Vandenabeele P, Droin N, Solary E
    Blood, 2011;118(17):4694-704.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  9. Design of protease-resistant myelin basic protein-derived peptides by cleavage site directed amino acid substitutions.
    Authors: Burster T, Marin-Esteban V, Boehm BO, Dunn S, Rotzschke O, Falk K, Weber E, Verhelst SH, Kalbacher H, Driessen C
    Biochem. Pharmacol., 2007;74(10):1514-23.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Enzyme Assay
  10. Cystatin M/E is a high affinity inhibitor of cathepsin V and cathepsin L by a reactive site that is distinct from the legumain-binding site. A novel clue for the role of cystatin M/E in epidermal cornification.
    Authors: Cheng T, Hitomi K, van Vlijmen-Willems IM, de Jongh GJ, Yamamoto K, Nishi K, Watts C, Reinheckel T, Schalkwijk J, Zeeuwen PL
    J. Biol. Chem., 2006;281(23):15893-9.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  11. Distinct roles for cysteine cathepsin genes in multistage tumorigenesis.
    Authors: Gocheva V, Zeng W, Ke D, Klimstra D, Reinheckel T, Peters C, Hanahan D, Joyce JA
    Genes Dev., 2006;20(5):543-56.
    Species: Human
    Sample Types: Buffer
    Applications: Enzyme Assay

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