Recombinant Human Cathepsin L Protein, CF Summary
Product Specifications
Glu113-Val333 & Ala114-Val333, both with a C-terminal 6-His tag
Analysis
Product Datasheets
952-CY
Carrier Free
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.
952-CY
Formulation | Supplied as a 0.2 μm filtered solution in Sodium Acetate 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.
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Assay Procedure
- Assay Buffer: 50 mM MES, 5 mM DTT, 1 mM EDTA, 0.005% (w/v) Brij-35, pH 6.0
- Recombinant Human Cathepsin L (rhCathepsin L) (Catalog # 952-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
- Dilute rhCathepsin L to 40 µg/mL in Assay Buffer.
- Incubate diluted rhCathepsin L on ice for 15 minutes.
- Dilute incubated 40 µg/mL rhCathepsin L to 0.02 ng/µL in Assay Buffer.
- Dilute Substrate to 80 µM in Assay Buffer.
- Load 50 µL of 0.02 ng/µL rhCathepsin L into a black well plate, and start the reaction by adding 50 µL of 80 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 80 µM Substrate without any rhCathepsin L.
- 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 L: 0.001 µg
- Substrate: 40 µM
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). Human Cathepsin L activity is greatest under mildly acidic conditions, from pH 4.5 - 6.5. The stability of the enzyme decreases at higher pH values.
- Goretzki, L. et al. (1992) FEBS Lett. 297:112.
- Taggart, C.C. et al. (2001) J. Biol. Chem. 276:33345.
- Gottesman, M.M. and F. Cabral (1981) Biochemistry 20:1659.
Citations for Recombinant Human 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.
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Citations: Showing 1 - 10
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The activation cascade of the broad-spectrum antiviral bemnifosbuvir characterized at atomic resolution
Authors: Chazot, A;Zimberger, C;Feracci, M;Moussa, A;Good, S;Sommadossi, JP;Alvarez, K;Ferron, F;Canard, B;
PLoS biology
Species: N/A
Sample Types: Recombinant Protein
Applications: Bioassay -
Pharmacologic inhibition of dipeptidyl peptidase 1 (cathepsin C) does not block in vitro granzyme-mediated target cell killing by CD8 T or NK cells
Authors: Sutton, VR;Watt, SV;Akhlaghi, H;Cipolla, DC;Chen, KJ;LaSala, D;McDonald, PP;Beavis, PA;Munoz, I;Hodel, AW;Noori, T;Voskoboinik, I;Trapani, JA;
Frontiers in pharmacology
Species: N/A
Sample Types: Recombinant Protein
Applications: Bioassay -
Autoantibody binding and unique enzyme-substrate intermediate conformation of human transglutaminase 3
Authors: Heggelund, JE;Das, S;Stamnaes, J;Iversen, R;Sollid, LM;
Nature communications
Species: N/A
Sample Types: Recombinant Protein
Applications: Bioassay -
mWTX-330, an IL 12 INDUKINE Molecule, Activates and Reshapes Tumor-infiltrating CD8+ T and NK Cells to Generate Antitumor Immunity
Authors: CJ Nirschl, HR Brodkin, C Domonkos, CJ Dwyer, DJ Hicklin, N Ismail, C Seidel-Dug, P Steiner, Z Steuert, JM Sullivan, WM Winston, A Salmeron
Cancer Immunology Research, 2023-04-19;0(0):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
mWTX-330, an IL 12 INDUKINE Molecule, Activates and Reshapes Tumor-infiltrating CD8+ T and NK Cells to Generate Antitumor Immunity
Authors: CJ Nirschl, HR Brodkin, C Domonkos, CJ Dwyer, DJ Hicklin, N Ismail, C Seidel-Dug, P Steiner, Z Steuert, JM Sullivan, WM Winston, A Salmeron
Cancer Immunology Research, 2023-04-19;0(0):.
Applications: Bioassay -
Hederacolchiside A1 Suppresses Autophagy by Inhibiting Cathepsin C and Reduces the Growth of Colon Cancer
Authors: S Kim, KH Lee, HJ Choi, E Kim, S Kang, M Han, HJ Jeon, MY Yun, GY Song, HJ Lee
Cancers, 2023-02-16;15(4):.
Species: N/A
Sample Types: Fluorogenic Peptide Substrate
Applications: Bioassay -
Cln5 represents a new type of cysteine-based S-depalmitoylase linked to neurodegeneration
Authors: AV Luebben, D Bender, S Becker, LM Crowther, I Erven, K Hofmann, J Söding, H Klemp, C Bellotti, A Stäuble, T Qiu, RS Kathayat, BC Dickinson, J Gärtner, GM Sheldrick, R Krätzner, R Steinfeld
Science Advances, 2022-04-15;8(15):eabj8633.
Species: N/A
Sample Types: Recombinant Protein
Applications: Bioassay -
A TMPRSS2 inhibitor acts as a pan-SARS-CoV-2 prophylactic and therapeutic
Authors: T Shapira, IA Monreal, SP Dion, DW Buchholz, B Imbiakha, AD Olmstead, M Jager, A Désilets, G Gao, M Martins, T Vandal, CAH Thompson, A Chin, WD Rees, T Steiner, IR Nabi, E Marsault, J Sahler, DG Diel, GR Van de Wal, A August, GR Whittaker, PL Boudreault, R Leduc, HC Aguilar, F Jean
Nature, 2022-03-28;0(0):.
Species: N/A
Sample Types: Fluorogenic Peptide Substrate
Applications: Bioassay -
Streptococcal pyrogenic exotoxin B cleaves GSDMA and triggers pyroptosis
Authors: W Deng, Y Bai, F Deng, Y Pan, S Mei, Z Zheng, R Min, Z Wu, W Li, R Miao, Z Zhang, TS Kupper, J Lieberman, X Liu
Nature, 2022-02-02;0(0):.
Species: Human
Sample Types: Recombinant Proteins
Applications: Bioassay -
Novel Cysteine Protease Inhibitor Derived from the Haementeria vizottoi Leech: Recombinant Expression, Purification, and Characterization
Authors: DDC Linhares, F Faria, RT Kodama, AMXP Amorim, FCV Portaro, D Trevisan-S, KF Ferraz, AM Chudzinski
Toxins, 2021-12-02;13(12):.
Species: Haementeria vizzotoi
Sample Types: Recombinant Protein
Applications: Bioassay -
Activation of Tenofovir Alafenamide and Sofosbuvir in the Human Lung and Its Implications in the Development of Nucleoside/Nucleotide Prodrugs for Treating SARS-CoV-2 Pulmonary Infection
Authors: J Li, S Liu, J Shi, HJ Zhu
Pharmaceutics, 2021-10-11;13(10):.
Species: Human
Sample Types: Whole Tissue
Applications: Enzyme Activity -
Decursin inhibits cell growth and autophagic flux in gastric cancer via suppression of cathepsin C
Authors: S Kim, SI Lee, N Kim, M Joo, KH Lee, MW Lee, HJ Jeon, H Ryu, JM Kim, JY Sul, GY Song, JY Kim, HJ Lee
American journal of cancer research, 2021-04-15;11(4):1304-1320.
Species: Human
Sample Types: Protein
Applications: Bioassay -
Direct Intracellular Visualization of Ebola Virus-Receptor Interaction by In Situ Proximity Ligation
Authors: E Mittler, T Alkutkar, RK Jangra, K Chandran
MBio, 2021-01-12;12(1):.
Species: Human
Sample Types: Protein
Applications: Bioassay -
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, 2019-09-29;20(19):.
Species: Human
Sample Types: Protein
Applications: Bioassay -
Cell penetrable, clickable and tagless activity-based probe of human cathepsin L
Authors: D Dana, J Garcia, AI Bhuiyan, P Rathod, L Joo, DA Novoa, S Paroly, KR Fath, EJ Chang, SK Pathak
Bioorg. Chem., 2019-02-14;85(0):505-514.
Species: Human
Sample Types: Recombinant Protein
Applications: Click Chemistry -
Image-guided surgery using near-infrared Turn-ON fluorescent nanoprobes for precise detection of tumor margins
Authors: R Blau, Y Epshtein, E Pisarevsky, G Tiram, S Israeli Da, E Yeini, A Krivitsky, A Eldar-Booc, D Ben-Shusha, H Gibori, A Scomparin, O Green, Y Ben-Nun, E Merquiol, H Doron, G Blum, N Erez, R Grossman, Z Ram, D Shabat, R Satchi-Fai
Theranostics, 2018-05-24;8(13):3437-3460.
Species: N/A
Sample Types: Polymer
Applications: Bioassay -
Suppression and resolution of autoimmune arthritis by rhesus ?-defensin-1, an immunomodulatory macrocyclic peptide
Authors: JB Schaal, DQ Tran, A Subramania, R Patel, T Laragione, KD Roberts, K Trinh, P Tongaonkar, PA Tran, D Minond, GB Fields, P Beringer, AJ Ouellette, PS Gulko, ME Selsted
PLoS ONE, 2017-11-16;12(11):e0187868.
Applications: Bioassay -
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, 2017-07-25;12(1):55.
Species: Human
Sample Types: Protein
Applications: Bioassay -
Immunization-Elicited Broadly Protective Antibody Reveals Ebolavirus Fusion Loop as a Site of Vulnerability
Authors: X Zhao, KA Howell, S He, JM Brannan, AZ Wec, E Davidson, HL Turner, CI Chiang, L Lei, JM Fels, H Vu, S Shulenin, AN Turonis, AI Kuehne, G Liu, M Ta, Y Wang, C Sundling, Y Xiao, JS Spence, BJ Doranz, FW Holtsberg, AB Ward, K Chandran, JM Dye, X Qiu, Y Li, MJ Aman
Cell, 2017-05-18;169(5):891-904.e15.
Species: Human
Sample Types: Complex Sample Type
Applications: Bioassay -
Enzymatic cleavage of myoferlin releases a dual C2-domain module linked to ERK signalling
Authors: AK Piper, SE Ross, GM Redpath, FA Lemckert, N Woolger, A Bournazos, PA Greer, RB Sutton, ST Cooper
Cell. Signal, 2017-02-10;33(0):30-40.
Species: Human
Sample Types: Protein
Applications: Bioassay -
Endosomal acidification and cathepsin L activity is required for calicivirus replication.
Authors: Shivanna V, Kim Y, Chang K
Virology, 2014-08-09;464(0):287-95.
Species: Porcine
Sample Types: Whole Cells
Applications: Bioassay -
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, 2014-06-04;5(0):3838.
Species: Mouse
Sample Types: Tissue Homogenates
Applications: Bioassay -
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, 2011-08-29;118(17):4694-704.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay -
Regulation of cathepsin g reduces the activation of proinsulin-reactive T cells from type 1 diabetes patients.
Authors: Zou F, Schafer N, Palesch D, Brucken R, Beck A, Sienczyk M, Kalbacher H, Sun Z, Boehm BO, Burster T
PLoS ONE, 2011-08-05;6(8):e22815.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay -
Development and validation of sandwich ELISA microarrays with minimal assay interference.
Authors: Gonzalez RM, Seurynck-Servoss SL, Crowley SA
J. Proteome Res., 2008-04-19;7(6):2406-14.
Applications: ELISA (Standard) -
Cystatin F is a cathepsin C-directed protease inhibitor regulated by proteolysis.
Authors: Hamilton G, Colbert JD, Schuettelkopf AW, Watts C
EMBO J., 2008-02-06;27(3):499-508.
Applications: Enzyme Assay -
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., 2007-08-02;74(10):1514-23.
Species: Human
Sample Types: Cell Lysates
Applications: Enzyme Assay -
Inhibition of the activation of multiple serine proteases with a cathepsin C inhibitor requires sustained exposure to prevent pro-enzyme processing.
Authors: Methot N, Rubin J, Guay D, Beaulieu C, Ethier D, Reddy TJ, Riendeau D, Percival MD
J. Biol. Chem., 2007-05-29;282(29):20836-46.
Applications: Enzyme Assay -
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., 2006-03-24;281(23):15893-9.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay -
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., 2006-02-15;20(5):543-56.
Species: Human
Sample Types: Buffer
Applications: Enzyme Assay
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Average Rating: 5 (Based on 2 Reviews)
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Reason for Rating: Works well for the kinase assay.
Reason for Rating: Excellent product. The consistency between lots seems to be high.