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| Source | Mouse Cathepsin B, amino acids (His18-Phe339) (Accession# P10605), with C-terminal 10x His tag, was expressed in CHO cells. |
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| Molecular Mass | The 332 amino acid recombinant protein has a predicted molecular mass of approximately 36.7 kD. The protein migrates at approximately 45 kD in DTT-reducing conditions and at approximately 40 kD in non-reducing conditions by SDS-PAGE. The predicted N-terminal amino acid is His. |
| Purity | >95%, as determined by Coomassie stained SDS-PAGE under reducing conditions. |
| Formulation | 0.22 µm filtered protein solution is in 25 mM Tris, 150 mM NaCl, and pH 7.5. |
| Endotoxin Level | Less than 0.01 ng per µg cytokine, as determined by the LAL method. |
| Concentration | 10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool. |
| Storage & Handling | Unopened vial can be stored at -20°C for one month, or at -70°C for six months. For maximum results, quick spin vial prior to opening. Avoid repeated freeze/thaw cycles. |
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| Activity |
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| The activity of Cathepsin B is determined by its ability to cleave the fluorogenic peptide substrate, Z-Leu-Arg-AMC (Z=Benzyloxycarbonyl, and AMC=7-amino-4-methylcoumarin) after activation. The specific activity is > 3600 pmol/min/µg. |
| Application | Bioassay |
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| Application Notes | Cathepsin B Activity Protocol: 1. Dilute the recombinant mouse Cathepsin B to 10 μg/ml in activation buffer (25 mM MES, 1 mM DTT, pH 5.0) and incubate the protein at 25°C for 3 min. Over activation may result in proteolysis and low activity. 2. Dilute the recombinant mouse Cathepsin B to 0.05 μg/ml in assay buffer (25 mM MES, pH 5.0). 3. Dilute the fluorogenic peptide substrate (Z-LR-AMC) to 40 μM using assay buffer. 4. Perform the assay in a 96-well, black, flat-bottomed plate. Add 50 μl of the diluted, activated Cathepsin B (0.0025 µg/well) to a black well and start the reaction by adding 50 μl of 40 μM substrate. The assay is read on a fluorometer at excitation and emission wavelengths of 380 nm and 460 nm for five minutes (top read). This protein is in the latent form and needs to be activated for bioassay. BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at [email protected]. |
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| Structure | Cysteine protease. |
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| Distribution | CTSB is ubiquitously expressed and is normally localized to lysosomes. However, it can also be secreted by many cells including tumor cells, macrophages, fibroblasts, osteoclasts, T lymphocytes, and neutrophils. |
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| Function | Cathepsin B is a lysosomal cysteine protease that is normally involved in intracellular protein turnover. |
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| Interaction | Interacts with Hepatitis B spliced protein, Bikunin, and TSRC1. |
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| Biology Area | Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Neurodegeneration, Neuroscience, Neuroscience Cell Markers, Protein Trafficking and Clearance, Stem Cells |
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| Molecular Family | Enzymes and Regulators, Lysosomal Markers |
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| Antigen References | 1. Mohamed MM, Sloane BF. 2006. Nat. Rev. Cancer. 6:764. 2. Wang C, et al. 2012. J. Biol. Chem. 47:39834. 3. Grzonka Z, et al. 2001. Acta Biochim. Pol. 48:1. 4. Liu J, et al. 2006. FEBS Letters 580:245. 5. Brix K, et al. 2008. Biochimie. 90:194. 6. Turk V, et al. 2012. Biochim. Biophys. Acta.1824:68. |
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| Gene ID | 13030 |
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| UniProt | View information about Cathepsin B on UniProt.org |
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| Regulatory Status | RUO |
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| Other Names | CTSB, CPSB, APP secretase, APPS, Cathepsin B1, CB |
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