Recombinant Mouse Cathepsin B Protein, CF
Recombinant Mouse Cathepsin B Protein, CF Summary
His18-Phe339, with a C-terminal 10-His tag
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.
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.
|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.
- 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
- Dilute rmCathepsin B to 10 µg/mL in Activation Buffer.
- Incubate at room temperature for 15 minutes (activation step).
- Dilute activated rmCathepsin B to 0.2 ng/µL in Assay Buffer.
- Dilute Substrate 20 µM in Assay Buffer.
- 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.
- 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:
- rmCathepsin B: 0.01 µg
- Substrate: 10 µM
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).
- Mort, J.S. (2004) in Handbook of Proteolytic Enzymes (Barrett, A.J. et al. eds.) p. 1079, Academic Press, San Diego.
- Vancompernolle, K. et al. (1998) FEBS Lett. 438:150.
- Jutras, I. and T.L. Reudelhuber (1998) FEBS Lett. 443:48.
- Taggart, C.C. et al. (2001) J. Biol. Chem. 276:33345.
- Berquin, I.M. and B.F. Sloane (1996) Adv. Exp. Med. Biol. 389:281.
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.
Citations: Showing 1 - 4
Filter your results:
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.
Sample Types: Whole Cells
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.
Sample Types: Whole Cells
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.
Sample Types: N/A
Applications: Binding Assay
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.
Sample Types: In Vivo
Applications: In Vivo
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Fluorogenic Peptide Substrates
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