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> Chemicals & Drugs > Amino Acid > Cathepsins

Cathepsins

Cathepsins are a family of lysosomal proteases found in most cell types.
They play crucial roles in intracellular protein degradation, antigen presentation, and tissue remodeling.
The Cathepsin family includes several subtypes, such as Cathepsin B, L, S, and K, each with distinct functions and substrate specificities.
These enzymes are involved in numerous physiological and pathological processes, including inflammation, cancer, and neurodegenerative disorders.
Researchers studying Cathepsins can optimize their work using PubCompare.ai, an AI-powered platform that enhances reproducibility and accuracy by helping locate protocols from literature, preprints, and patents, while leveraging AI-driven comparisons to identify the best protocols and products.
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Most cited protocols related to «Cathepsins»

1
Gelatin Zymography for Cathepsin Activity
Cited 20 times

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Publication 2011
Acetic Acid Biological Assay Bromphenol Blue Buffers Cathepsins Cells Centrifugation Coomassie blue Edetic Acid Egtazic Acid Elastin Electrophoresis Enzymes Gelatins Gels Glycerin Glycerophosphates Homo sapiens Isopropyl Alcohol leupeptin Orthovanadate polyacrylamide gels Proteins Sodium Sodium Acetate Sodium Chloride sodium phosphate Stains Tissues Triton X-100 Tromethamine Tween 20
2
Phagosome Isolation and Protein Analysis
Cited 18 times
Detailed descriptions of reagents and experiments can be found in Supplementary Methods. All stable cell lines were generated by retroviral gene transfer. For phagosome isolation, RAW cells were fed latex beads (Polysciences), the cells were disrupted by Dounce homogenization, and latex bead containing phagosomes were purified on a 62%−10% sucrose step gradient in a Beckman SW40Ti centrifuge. After purification, phagosomes were lysed by addition of TritonX-100, and proteins in phagosomes were separated by SDS-PAGE and visualized by immunoblot. Pulse/chase analyses were performed by starving cells of cysteine/methionine for 1 hour, pulsing with 35S-cysteine/methionine for 45 minutes, and chasing in molar excess of unlabeled cysteine/methionine for the indicated time periods. After lysis, radiolabeled proteins were immunoprecipated and visualized by SDS-PAGE. Deglycosylation assays were performed on immunoprecipitated proteins using endoglycosidaseH or PNGaseF (both from NEB) with supplied buffers according to manufacturer's instructions. Proteins were separated by SDS-PAGE and visualized by immunoblot. In vitro proteolysis with recombinant cathepsins (Biomol) was performed according to manufacturer guidelines. For MyD88/TLR9 coimmunoprecipitations, cells were lysed in RIPA Buffer before or after stimulation with CpG, and protein complexes were precipitated with anti-FLAG matrix. For CpG binding assays, TLR9-HA containing lysates were incubated with Biotin-CpG followed by precipitation with streptavidin matrix. In both cases, precipitated proteins were visualized by SDS-PAGE followed by immunoblot. NF-κB luciferase assays were performed by transient transfection of HEK293T cells with an NF-κB luciferase reporter plasmid and the indicated expression plasmids using LTX transfection reagent (Invitrogen) according to manufacturer's instructions. C57/Bl/6 mice were purchased from Jackson Laboratories. All knockout mice have been previously described19 (link)-22 (link). Mice were housed within animal facilities at UC Berkeley or UCSF according to IACUC guidelines.
Ewald S.E., Lee B.L., Lau L., Wickliffe K.E., Shi G.P., Chapman H.A, & Barton G.M. (2008). The ectodomain of Toll-like receptor 9 is cleaved to generate a functional receptor. Nature, 456(7222), 658-662.
Publication 2008
Animals Biological Assay Biotin Buffers Cathepsins Cell Lines Cells Cysteine Gene Transfer, Horizontal Immunoblotting Institutional Animal Care and Use Committees isolation Luciferases Methionine Mice, Inbred C57BL Mice, Knockout Molar Mus Phagosomes Plasmids Proteins Proteolysis Radioimmunoprecipitation Assay RELA protein, human Retroviridae SDS-PAGE Streptavidin Sucrose Transfection Transients
3
Phagosome Isolation and Protein Analysis
Cited 18 times
Detailed descriptions of reagents and experiments can be found in Supplementary Methods. All stable cell lines were generated by retroviral gene transfer. For phagosome isolation, RAW cells were fed latex beads (Polysciences), the cells were disrupted by Dounce homogenization, and latex bead containing phagosomes were purified on a 62%−10% sucrose step gradient in a Beckman SW40Ti centrifuge. After purification, phagosomes were lysed by addition of TritonX-100, and proteins in phagosomes were separated by SDS-PAGE and visualized by immunoblot. Pulse/chase analyses were performed by starving cells of cysteine/methionine for 1 hour, pulsing with 35S-cysteine/methionine for 45 minutes, and chasing in molar excess of unlabeled cysteine/methionine for the indicated time periods. After lysis, radiolabeled proteins were immunoprecipated and visualized by SDS-PAGE. Deglycosylation assays were performed on immunoprecipitated proteins using endoglycosidaseH or PNGaseF (both from NEB) with supplied buffers according to manufacturer's instructions. Proteins were separated by SDS-PAGE and visualized by immunoblot. In vitro proteolysis with recombinant cathepsins (Biomol) was performed according to manufacturer guidelines. For MyD88/TLR9 coimmunoprecipitations, cells were lysed in RIPA Buffer before or after stimulation with CpG, and protein complexes were precipitated with anti-FLAG matrix. For CpG binding assays, TLR9-HA containing lysates were incubated with Biotin-CpG followed by precipitation with streptavidin matrix. In both cases, precipitated proteins were visualized by SDS-PAGE followed by immunoblot. NF-κB luciferase assays were performed by transient transfection of HEK293T cells with an NF-κB luciferase reporter plasmid and the indicated expression plasmids using LTX transfection reagent (Invitrogen) according to manufacturer's instructions. C57/Bl/6 mice were purchased from Jackson Laboratories. All knockout mice have been previously described19 (link)-22 (link). Mice were housed within animal facilities at UC Berkeley or UCSF according to IACUC guidelines.
Ewald S.E., Lee B.L., Lau L., Wickliffe K.E., Shi G.P., Chapman H.A, & Barton G.M. (2008). The ectodomain of Toll-like receptor 9 is cleaved to generate a functional receptor. Nature, 456(7222), 658-662.
Publication 2008
Animals Biological Assay Biotin Buffers Cathepsins Cell Lines Cells Cysteine Gene Transfer, Horizontal Immunoblotting Institutional Animal Care and Use Committees isolation Luciferases Methionine Mice, Inbred C57BL Mice, Knockout Molar Mus Phagosomes Plasmids Proteins Proteolysis Radioimmunoprecipitation Assay RELA protein, human Retroviridae SDS-PAGE Streptavidin Sucrose Transfection Transients
4
Lysosomal Membrane Permeabilization Assay
Cited 15 times

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Publication 2015
Caspase Cathepsins Cell Extracts Cells Cholesterol Cysteine Cytosol Detergents Digitonin Glucosaminidases Hydrolase Lysosomes Plasma Membrane Tissue, Membrane Titrimetry
5
Affinity Purification of Protein Antibodies
Cited 14 times
The monoclonal anti-CD8 was obtained from Qbiogene. Anti-GM130 and anti-EEA1 were purchased from Becton Dickinson, and the anti-TfnR antibody was obtained from Zymed Laboratories. Polyclonal anti-actin was purchased from Sigma-Aldrich, and the polyclonal anti-CI-MPR (Reaves et al., 1996 (link)) was a gift from J. Paul Luzio (University of Cambridge, Cambridge, UK). Anti-cathepsin D antiserum was obtained from DakoCytomation, and anti-TGN46 was supplied by Serotec. Polyclonal anti-mVPS26, anti-Snx1, and anti-mVPS35 were produced in house using GST fusion proteins as antigens. The antisera were subsequently affinity purified using the respective antigens coupled to CNBr-Sepharose.
, & Seaman M.N. (2004). Cargo-selective endosomal sorting for retrieval to the Golgi requires retromer. The Journal of Cell Biology, 165(1), 111-122.
Publication 2004
Actins anti-d antibody Antibodies, Anti-Idiotypic Antigens BDP1 protein, human Cathepsins Cyanogen Bromide IGF2R protein, human Immune Sera Proteins Sepharose

Most recents protocols related to «Cathepsins»

1
Measuring Cathepsin K Protease Activity
The protease activity of cathepsin K from cultured media of OCs was measured using a Cathepsin K Activity Assay Kit (Abcam) in accordance with the manufacturer’s protocol.
Kim E.Y., Kim J.E., Kim Y.E., Choi B., Sohn D.H., Park S.O., Chung Y.H., Kim Y., Robinson W.H., Kim Y.G, & Chang E.J. (2023). Dysfunction in parkin aggravates inflammatory bone erosion by reinforcing osteoclast activity. Cell & Bioscience, 13, 48.
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Publication 2023
Biological Assay Cathepsins CTSK protein, human Endopeptidase K Peptide Hydrolases
2
Quantification of Chitinase and Cathepsin in Infected Cells
Chitinase and cathepsin levels in infected SF-21 cells were measured at 48 h p.i. in three independent experiments, as described earlier [19 (link)]. Mock- and virus-infected cells (48 h p.i.) were assessed for chitinase activity, using 50 μg total protein and CM-chitin-RBV substrate (Loewe Biochemica GmbH, Sauerlach, Bayern, Germany) [19 (link)], and cysteine protease activity, using 400 μg total protein and azocasein substrate, in the presence and absence of a cysteine protease inhibitor (E-64, 20 μM), based on a previously described assay [15 (link)]. Enzyme assays of infected insect cadavers were similar but 200 μg or 400 μg of total homogenate protein was used for the CHIA or V-CATH assays, respectively. T-tests (unpaired, two-tailed, 95% CI) were performed using GraphPad Prism 9.2.0.
Hodgson J.J., Passarelli A.L, & Krell P.J. (2023). Transcriptional Reprogramming of Autographa Californica Multiple Nucleopolyhedrovirus Chitinase and Cathepsin Genes Enhances Virulence. Viruses, 15(2), 503.
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Publication 2023
azocasein Biological Assay Cadaver Cathepsins Cells Chitinases Cysteine Proteases Cysteine Proteinase Inhibitors Enzyme Assays Insecta O-(carboxymethyl)chitin prisma Proteins Salvia hispanica seed Virus
3
Subcellular Tracking of Cathepsin D and Mitophagy
After CTL treatment for 24 h, breast cancer cells were fixed with 4% PFA in PBS and permeabilized with 0.5% Triton X-100 for 7 min. After PBS washing, cells were blocked with PBS-T containing 10% FBS and 1% BSA for 1 h. To measure the release of cathepsin D into the cytosol, cells were incubated with anti-cathepsin D primary antibody (sc-6486, 1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA) overnight and followed by Alexa Flour 488-conjugated anti-goat IgG for 1 h (1:250, Invitrogen, Carlsbad, CA, USA). Nuclei were stained with 1 μg/mL DAPI (Sigma, St. Louis, MO, USA) in 2% BSA for 1 min. To analyze the mitophagy induction through the interaction of mitochondria with the lysosome, cells were incubated with primary antibodies (anti-COX Ⅳ, #4850, 1:250, Cell Signaling, Danvers, MA, USA and anti-LAMP2, sc-18822, 1:250, Santa Cruz Biotechnology, Santa Cruz, CA, USA). Cells were incubated with Alexa Flour 488-conjugated anti-rabbit IgG and 594-conjugated anti-mouse IgG (1:250, Invitrogen, Carlsbad, CA, USA) for 1 h. The autophagosome-mediated mitophagy was analyzed by staining with anti-COX Ⅳ in GFP-LC3-transfected cells, followed by incubating with Alexa Flour 594-conjugated anti-rabbit IgG. To assess the integrity of lysosome, cells were treated with CTL for 24 h and stained with 50 nM Lysotracker Red DND-99 at 37 °C for 30 min. Fluorescence images were acquired using confocal microscopy (Carl Zeiss, Jena, Germany).
Choi Y.J., Choi Y.K., Ko S.G., Cheon C, & Kim T.Y. (2023). Investigation of Molecular Mechanisms Involved in Sensitivity to the Anti-Cancer Activity of Costunolide in Breast Cancer Cells. International Journal of Molecular Sciences, 24(4), 4009.
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Publication 2023
Aftercare Alexa594 anti-d antibody anti-IgG Antibodies Autophagosome Breast Carcinoma Cathepsin D Cathepsins Cell Nucleus Cells Cytosol DAPI Flour Fluorescence Goat LAMP2 protein, human Lysosomes LysoTracker Microscopy, Confocal Mitochondria Mitophagy Mus Rabbits Red DND-99 Triton X-100
4
Cathepsin B Immunofluorescence in db/db Mice
The paraffin-embedded tissue sections from vehicle or metformin treated db/db mice were used for staining. After deparaffinization (Xylene (twice, 5 min), 100% ethanol (twice, 5 min), 95% ethanol (twice, 5 min), 70% ethanol (once, 5 min), 50% ethanol (once, 5 min), and type one water, the tissue was boiled in 10 mM sodium citrate buffer (Vector labs) for 20 min. After a PBS wash, blocking was performed with 2.5% normal horse serum (Vector labs) at room temperature. Next, the tissue sections were incubated with cathepsin B antibody ((1:500) Cell Signaling Tech, 3383) for 2 h and incubated with a fluoresceine-labeled horse anti-rabbit secondary for 1 hr. After washing with PBS, slides were mounted using Vectashield anti-fade mounting media. Fluorescence images were captured using an Eclipse Ti2 by Nikon Instruments Inc., Melville, NY, USA.
Scindia Y.M., Gholam M.F., Waleed A., Liu L.P., Chacko K.M., Desai D., Lopez J.P., Malik Z., Schramm W.C., Morales A.G., Carson-Marino M, & Alli A.A. (2023). Metformin Alleviates Diabetes-Associated Hypertension by Attenuating the Renal Epithelial Sodium Channel. Biomedicines, 11(2), 305.
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Publication 2023
Buffers Cathepsins Cloning Vectors CTSB protein, human Equus caballus Ethanol Fluorescein Fluorescence immunoglobulin B Immunoglobulins Metformin Mus Paraffin Embedding Rabbits Serum Sodium Citrate Tissues Xylene
5
Labeling Cysteine Cathepsins in Cells
Cysteine cathepsins can be labelled in cultured cells by using the activity-based probe DCG04 (Medkoo, Morrisville, NC, USA), which is a biotinylated form of general cysteine peptidase inhibitor E-64. DCG04 is a selective activity-based probe of cysteine peptidases. It is a biotinylated form of the general cysteine peptidase inhibitor E64 (which line number). It has the ability to bind to active cysteine peptidases in protein mixture [37 (link)]. Cells were incubated for 72 h at 37 °C with 10 μM DCG04. Protein extract was then prepared. 30 μg of the extract were separated using 12.5% SDS polyacrylamide gels. Proteins were then transferred to nitrocellulose membrane (Santa Cruz biotechnology, Dallas, TX, USA). Membranes were then blocked using 3% bovine serum albumin (BSA) (ThermoFischer Scientific, Waltham, MA, USA) in PBS. Membrane was then incubated in streptavidin-horseraddish peroxidase (0.125 μg/mL PBST (BioLegend, San Diego, CA, USA) prior to enhanced chemiluminescence detection.
Aufy M., Abdelaziz R.F., Hussein A.M., Topcagic N., Shamroukh H., Abdel-Maksoud M.A., Salem T.Z, & Studenik C.R. (2023). Impact of Enniatin B and Beauvericin on Lysosomal Cathepsin B Secretion and Apoptosis Induction. International Journal of Molecular Sciences, 24(3), 2030.
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Publication 2023
Cathepsins Cells Chemiluminescence Cultured Cells Cysteine Cysteine Proteases Nitrocellulose Peroxidase polyacrylamide gels Proteins Serum Albumin, Bovine Streptavidin Tissue, Membrane

Top products related to «Cathepsins»

1
Anti cathepsin d by Santa Cruz Biotechnology
Sourced in United States
Anti-cathepsin D is a laboratory reagent used to detect and quantify the presence of cathepsin D, a lysosomal aspartic protease, in biological samples. It can be used in various research applications, such as cell biology, molecular biology, and biochemistry.
2
Prosense 680 by PerkinElmer
Sourced in United States, Germany, Morocco
ProSense 680 is a sensitive and versatile fluorescence detector designed for a wide range of life science applications. It offers high-performance detection capabilities with a simple and intuitive user interface.
3
Pvdf membrane by Merck Group
Sourced in United States, Germany, China, United Kingdom, Morocco, Ireland, France, Italy, Japan, Canada, Spain, Switzerland, New Zealand, India, Hong Kong, Sao Tome and Principe, Sweden, Netherlands, Australia, Belgium, Austria
PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.
4
Anti cathepsin d by Abcam
Sourced in United States
Anti-cathepsin D is a laboratory reagent used to detect and quantify the presence of cathepsin D, an aspartic protease enzyme, in various biological samples. It can be utilized in techniques such as Western blotting, immunohistochemistry, and ELISA to analyze cathepsin D levels without interpreting its intended use.
5
Chloroquine by Merck Group
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Chloroquine is a laboratory chemical primarily used as a research tool in biochemical and cell biology applications. It is a white, crystalline solid that is soluble in water. Chloroquine is commonly used in experiments to study cellular processes, such as autophagy and endocytosis, by inhibiting the function of lysosomes. Its core function is to serve as a research reagent for scientific investigations, without making any claims about its intended use.
6
Bafilomycin a1 by Merck Group
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Bafilomycin A1 is a macrolide compound that acts as a potent and specific inhibitor of vacuolar-type H+-ATPases (V-ATPases). V-ATPases are involved in the acidification of various intracellular compartments, making Bafilomycin A1 a useful tool for studying cellular processes that rely on pH regulation.
7
Anti cathepsin d by Cell Signaling Technology
Sourced in United States
Anti-Cathepsin D is a primary antibody that specifically recognizes the Cathepsin D protein. Cathepsin D is an aspartic protease involved in intracellular protein breakdown and tissue remodeling.
8
Protease inhibitor cocktail by Merck Group
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The Protease Inhibitor Cocktail is a laboratory product designed to inhibit the activity of proteases, which are enzymes that can degrade proteins. It is a combination of various chemical compounds that work to prevent the breakdown of proteins in biological samples, allowing for more accurate analysis and preservation of protein integrity.
9
Lysotracker red dnd 99 by Thermo Fisher Scientific
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LysoTracker Red DND-99 is a fluorescent dye that selectively stains acidic organelles, such as lysosomes, in live cells. It can be used to visualize and monitor the distribution and activity of lysosomes within the cellular environment.
10
Cathepsin b by Merck Group
Sourced in United States, United Kingdom
Cathepsin B is a laboratory equipment product manufactured by Merck Group. It is a proteolytic enzyme that plays a role in various cellular processes. The core function of Cathepsin B is to cleave peptide bonds within proteins.

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