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Recombinant Mouse Cathepsin L Protein, CF

Catalog #: 1515-CY
8 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
1515-CY-010
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Citations (8)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Mouse Cathepsin L Protein, CF Summary

Product Specifications

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 >5,000 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived mouse Cathepsin L protein
Thr18-Asn334, with a C-terminal 10-His tag
Accession #
P06797
N-terminal Sequence
Analysis
Thr18
Structure / Form
Pro form
Predicted Molecular Mass
37 kDa
SDS-PAGE
35-40 kDa, reducing conditions

Product Datasheets

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1515-CY

Product Datasheet
COA
SDS

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.

1515-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: 50 mM Sodium Citrate, 150 mM NaCl, 1 mM EDTA, 0.615% CHAPS, pH 3.0
  • Assay Buffer: 25 mM MES, 5 mM DTT, pH 6.0
  • Recombinant Mouse Cathepsin L (rmCathepsin L) (Catalog # 1515-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. Activate rmCathepsin L at 100 µg/mL in Activation Buffer.
  2. Incubate at room temperature for 60 minutes.
  3. Dilute activated rmCathepsin L to 0.05 ng/µL in Assay Buffer.
  4. Dilute Substrate to 20 µM in Assay Buffer.
  5. In a plate load 50 µL of 0.05 ng/µL rmCathepsin L to wells, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 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 L: 0.0025 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 L

Cathepsin L is a lysosomal cysteine protease expressed in most eukaryotic cells. Cathepsin L is known to hydrolyze a number of proteins, including the proform of urokinase‑type plasminogen activator, which is activated by Cathepsin L cleavage (1). Cathepsin L has also been shown to proteolytically inactivate alpha 1-antitrypsin and secretory leucoprotease inhibitor, two major protease inhibitors of the respiratory tract (2). These observations, combined with the demonstration of increased Cathepsin L activity in the epithelial lining fluid of the lungs of emphysema patients, have led to the suggestion that the enzyme may be involved in the progression of this disease. Cathepsin L has also been identified as a major excreted protein of transformed fibroblasts, indicating the enzyme could be involved in malignant tumor growth (3). In Cathepsin L‑deficient mice, it appears to play a critical role in cardiac morphology and function, epidermal homeostasis, regulation of the hair cycle, and MHC class II‑mediated antigen presentation in cortical epithelial cells of the thymus (4, 5). Mouse Cathepsin L is synthesized as a 334 amino acid precursor with a signal peptide (residues 1‑17), a pro region (residues 18‑113) and a mature chain (residues 114‑334).

References
  1. Goretzki, L. et al. (1992) FEBS Lett. 297:112.
  2. Taggart, C.C. et al. (2001) J. Biol. Chem. 276:33345.
  3. Gottesman, M.M. and F. Cabral (1981) Biochemistry 20:1659.
  4. Stypmann, J. et al. (2002) Proc. Natl. Acad. Sci. USA 99: 6234.
  5. Reinheckel, T. et al. (2001) Biol. Chem. 382:735.
    6.    
Entrez Gene IDs
1514 (Human); 13039 (Mouse); 118276442 (Insect)
Alternate Names
Cathepsin L; cathepsin L1; CATL; CTSL; CTSL1; CTSLEC 3.4.22.15; EC 3.4.22; FLJ31037; Major excreted protein; MEP

Citations for Recombinant Mouse Cathepsin L 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.

8 Citations: Showing 1 - 8
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  1. Effect of cysteine peptidase inhibitor of Eudiplozoon nipponicum (Monogenea) on cytokine expression of macrophages in vitro
    Authors: J Ilgová, L Kavanová, K Matiašková, J Salát, M Kašný
    Mol. Biochem. Parasitol., 2019-12-23;235(0):111248.
    Species: Diplozoidae - Eudiplozoon nipponicum
    Sample Types: Recombinant Protein
    Applications: Bioassay
  2. A novel type I cystatin of parasite origin with atypical legumain-binding domain
    Authors: J Ilgová, L Jedli?ková, H Dvo?áková, M Benovics, L Mikeš, L Janda, J Vorel, P Roudnický, D Pot?šil, Z Zdráhal, M Gelnar, M Kašný
    Sci Rep, 2017-12-13;7(1):17526.
    Species: Mouse
    Sample Types: Recombinant Protein
    Applications: Bioassay
  3. Extracellular cathepsin L stimulates axonal growth in neurons
    Authors: C Tohda, M Tohda
    BMC Res Notes, 2017-11-23;10(1):613.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  4. Lysosomal processing of progranulin
    Authors: X Zhou, DH Paushter, T Feng, L Sun, T Reinheckel, F Hu
    Mol Neurodegener, 2017-08-23;12(1):62.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  5. Supraspinatus tendon overuse results in degenerative changes to tendon insertion region and adjacent humeral cartilage in a rat model
    Authors: Akia N Parks
    J. Orthop. Res, 2016-12-21;0(0):.
    Applications: Bioassay
  6. Cathepsins limit macrophage necroptosis through cleavage of Rip1 kinase.
    Authors: McComb S, Shutinoski B, Thurston S, Cessford E, Kumar K, Sad S
    J Immunol, 2014-05-05;192(12):5671-8.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  7. The cystatin M/E-cathepsin L balance is essential for tissue homeostasis in epidermis, hair follicles, and cornea.
    Authors: Zeeuwen PL, van Vlijmen-Willems IM, Cheng T
    FASEB J., 2010-05-21;24(10):3744-55.
    Species: Mouse
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  8. Cathepsin L proteolytically processes histone H3 during mouse embryonic stem cell differentiation.
    Authors: Duncan EM, Muratore-Schroeder TL, Cook RG, Garcia BA, Shabanowitz J, Hunt DF, Allis CD
    Cell, 2008-10-17;135(2):284-94.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay

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