Log In Sign up
The largest database of trusted experimental protocols
> Chemicals & Drugs > Amino Acid > Calpeptin

Calpeptin

Calpeptin is a a cell-permeable, non-peptide calpain inhibitor that has been widely used in biological research to investigate the roles of calpain proteases.
It functions by selectively and reversibly inhibiting the activity of calpain, a calcium-dependent cysteine protease involved in various cellular processes, such as cytoskeletal remodeling, signal transduction, and apoptosis.
Calpeptin has been utilized in studies to elucidate the involvment of calpain in physiological and pathological conditions, includeing neurodegeneration, cancer, and inflammation.
Resesarchers can leverage PubCompare.ai's AI-driven platform to easily identify and compare the most relevant Calpeptin research protocols from the literature, preprints, and patents, optimizing their investigations into this important protease inhibitor.

Most cited protocols related to «Calpeptin»

1
Quantifying Activation of Rho GTPases
Cited 7 times
Swiss 3T3 cells were continuously passaged at subconfluence and used to monitor the capacity of compounds to block stimulus-mediated activation of Cdc42 or Rac1 in vivo. Vero E6 cells are responsive to calpeptin-mediated stimulation of RhoA and thus were used as a second cell line for assessing compound effects on RhoA activity. Commercial GLISA kits customized to capture activated Cdc42, Rac1, or RhoA from cell lysates were used per manufacturer's instructions (Cytoskeleton). Cells were serum-starved by sequentially removing serum over a 3-day period and treated with either 0.1% DMSO or 1 or 10 μm compound for 60 min. Subsequently, samples were treated with 100 ng/ml EGF for 2 min to activate Cdc42 or Rac1 or with 1 μg/ml calpeptin for 30 min to activate RhoA. DMSO-treated cells were left either unstimulated to determine base-line GTPase activation or stimulated without drug treatment to determine maximal GTPase activation. Cell lysates were frozen, and active Cdc42 and Rac1 were quantified based on a p21-activated protein kinase binding assay (27 (link)), whereas active RhoA was quantified based on a rhotekin binding assay (28 (link)). All assays were performed in 96-well microtiter plates. Purified GTP-loaded Cdc42, Rac1, or RhoA control proteins were included in each assay to quantify active GTPases in the lysates.
RhoA activation was independently confirmed by monitoring actin stress fiber formation following calpeptin stimulation (1 μg/ml) for 30 min. Actin filaments in control and stimulated cells were detected following paraformaldehyde (4%) fixation and Triton X-100 (0.5%) permeabilization using Alexa Fluor® 488 phalloidin (1 unit, or ∼0.17 μm) (Life Technologies).
Hong L., Kenney S.R., Phillips G.K., Simpson D., Schroeder C.E., Nöth J., Romero E., Swanson S., Waller A., Strouse J.J., Carter M., Chigaev A., Ursu O., Oprea T., Hjelle B., Golden J.E., Aubé J., Hudson L.G., Buranda T., Sklar L.A, & Wandinger-Ness A. (2013). Characterization of a Cdc42 Protein Inhibitor and Its Use as a Molecular Probe. The Journal of Biological Chemistry, 288(12), 8531-8543.
Publication 2013
Actins alexa fluor 488 BaseLine dental cement Biological Assay calpeptin Cardiac Arrest CDC42 protein, human Cell Lines Cells Cytoskeleton Freezing Guanosine Triphosphate Phosphohydrolases Microfilaments p21-Activated Kinases paraform Phalloidine Pharmaceutical Preparations Proteins rhoA GTP-Binding Protein RHOA protein, human rhotekin protein, human Serum Stress Fibers Sulfoxide, Dimethyl Swiss 3T3 Cells Triton X-100 Vero Cells
2
Mechanical Wounding and Cell Survival Assay
Cited 4 times
MLO-Y4, OCY454, and primary osteocytes were wounded by glass beads, as previously described (8 (link), 19 (link)), to facilitate wounding large numbers of cells. Vitamin E (Sigma T3251, 220 μM) (19 (link)) or calpeptin (Sigma C8999, 20 μM) (20 (link)) were introduced as indicated 24 hours prior to wounding. Whole cell lysates were collected for western blotting as previously described (21 ), using antibodies against c-fos (Santa Cruz) and β-actin (Sigma). Please see Supplementary Methods for additional details.
To assess cell survival after mechanical wounding, MLO-Y4 cells were wounded by glass beads in the presence of lysine-fixable fluorescein-conjugated dextran (10 kDa, 5 mg/mL+10 mg/mL BSA) containing either 1.8 mM Ca2+, 1.8 mM Ca2++20 μM calpeptin, or 1.5 mM EGTA. Five minutes after wounding, cells were stained with propidium iodide (0.3 μg/mL) to detect dead cells (i.e., unrepaired PMD) and imaged on a multi-photon confocal microscope (Zeiss). Please see Supplementary Methods for additional details. The percentages of PMD-affected cells and dead cells were quantified (Bioquant).
Yu K., Sellman D.P., Bahraini A., Hagan M.L., Elsherbini A., Vanpelt K.T., Marshall P.L., Hamrick M.W., McNeil A., McNeil P.L, & McGee-Lawrence M.E. (2017). Mechanical loading disrupts osteocyte plasma membranes which initiates mechanosensation events in bone. Journal of orthopaedic research : official publication of the Orthopaedic Research Society, 36(2), 653-662.
Publication 2017
Actins Antibodies calpeptin Cells Cell Survival Egtazic Acid fluorescein-dextran Lysine Microscopy, Confocal Osteocytes Propidium Iodide Vitamin E
3
Synthesis and Characterization of PU24FCl, PU-H71, YK5, and YK5B
Cited 3 times

Protocol full text hidden due to copyright restrictions

Open the protocol to access the free full text link

Publication 2013
4-(2-aminoethyl)benzenesulfonylfluoride Anabolism benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone calpeptin Cathepsins leupeptin MG 132 PU-H71
4
Purification of Drp1 and Actin for Biochemical Studies
Cited 3 times
Drp1 was expressed in One Shot BL21 Star (DE3) Escherichia coli (C6010-03; Life Technologies, Carlsbad, CA) by isopropyl-β-d-thiogalactoside induction at 20°C for 16 h. Cell pellets were resuspended in lysis buffer (100 mM Tris-Cl, pH 8.0, 500 mM NaCl, 1 mM dithiothreitol [DTT], 1 mM EDTA, 2 μg/ml leupeptin, 10 μg/ml aprotinin, 2 μg/ml pepstatin A, 2 mM benzamidine, 1 μg/ml calpain inhibitor I [ALLN], and 1 μg/ml calpeptin) and lysed using a high-pressure homogenizer (M-110L Microfluidizer Processor; Microfluidics, Newton, MA). The lysate was clarified by centrifugation at 40,000 rpm (type 45 Ti rotor; Beckman, Brea, CA) for 1 h at 4°C. Avidin (20 μg/ml; PI-21128; ThermoFisher Scientific, Waltham, MA) was added, and then the supernatant was loaded onto Strep-Tactin Superflow resin (2-1206-025; IBA, Göttingen, Germany) by gravity flow. The column was washed with 20-column volumes of lysis buffer without protease inhibitors. To elute Drp1, 0.01 mg/ml HRV3C protease in lysis buffer without protease inhibitors was added for 16 h at 4°C. The Strep-Tactin Superflow eluate was further purified by size exclusion chromatography on Superdex200 (GE Biosciences, Piscataway, NJ), spin concentrated, frozen in liquid nitrogen, and stored at −80°C.
Rabbit skeletal muscle actin was extracted from acetone powder as previously described (Spudich and Watt, 1971 (link)), and further purified by size exclusion chromatography on Superdex 75 (GE Biosciences). Actin was stored at 4°C in G-buffer (2 mM Tris, pH 8.0, 0.5 mM DTT, 0.2 mM ATP, 0.1 mM CaCl2, and 0.01% NaN3).
Hatch A.L., Ji W.K., Merrill R.A., Strack S, & Higgs H.N. (2016). Actin filaments as dynamic reservoirs for Drp1 recruitment. Molecular Biology of the Cell, 27(20), 3109-3121.
Publication 2016
Acetone acetylleucyl-leucyl-norleucinal Actins Aprotinin Avidin benzamidine Buffers calpeptin Cells Centrifugation Dithiothreitol Edetic Acid Endopeptidases Escherichia coli Freezing Gel Chromatography Gravity leupeptin Nitrogen Pellets, Drug pepstatin Powder Pressure Protease Inhibitors Rabbits Resins, Plant Skeletal Muscles Sodium Azide Sodium Chloride Streptococcal Infections Tromethamine
5
Modulating TRPM6/7 sensitivity in colorectal cancer
Cited 3 times
Colon cancer LoVo cells sensitive (LoVo-S) or resistant to DXR (LoVo-R) (kindly donated by Dr. P. Perego, Istituto Nazionale Tumori, Milano) were cultured in DMEM containing 10% fetal bovine serum and 2 mM glutamine at 37 °C and 5% CO2.
To obtain a transient downregulation of TRPM6 and 7, we utilized the stealth siRNAs developed by Qiagen37 (link). siRNAs were transfected into 2 × 104/cm2 cells using HiPerFect Transfection Reagent (Qiagen). Non-silencing, scrambled sequences were used as controls (CTR).
To evaluate sensitivity to DXR, cell viability was assessed by MTT assay. Briefly, cells were seeded in 96 well/plates. Where indicated, 16 h prior to DXR treatment, LoVo-S were silenced for TRPM7 or TRPM6, or exposed to 2-aminoethoxydiphenyl borate (2-APB, 50 μM), while LoVo-R were treated with calpeptin (5 μg/ml). After 48 h of DXR treatment (1 μg/ml), culture medium was replaced with medium containing 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT, 0.5 mg/ml) (Sigma, Oakville, Ontario, Canada). At the end of the experiment, media were removed and formazan crystals generated by cellular reductase activity were dissolved in DMSO:ethanol (1:1) and quantified by absorbance readings at 575 nm. The experiment was repeated three times in quadruplicates. Percentage cell viability was calculated from the absorbance measured in DXR-treated vs. untreated cells for each experimental condition.
To assess DXR retention, cells were loaded with 10 μM DXR for 2 h and fixed either immediately (loading control) or 30 min after washing (efflux). Nuclear fluorescence was visualized by confocal microscopy and quantified as the mean nuclear fluorescence measured in efflux conditions vs. loading control.
Castiglioni S., Cazzaniga A., Trapani V., Cappadone C., Farruggia G., Merolle L., Wolf F.I., Iotti S, & Maier J.A. (2015). Magnesium homeostasis in colon carcinoma LoVo cells sensitive or resistant to doxorubicin. Scientific Reports, 5, 16538.
Full text: Click here
Publication 2015
Aftercare Biological Assay Borates Bromides calpeptin Cancer of Colon Cells Cell Survival Down-Regulation Ethanol Fetal Bovine Serum Fluorescence Formazans Glutamine Hypersensitivity Microscopy, Confocal Oxidoreductase Retention (Psychology) RNA, Small Interfering Sulfoxide, Dimethyl Training Programs Transfection Transients TRPM6 protein, human TRPM7 protein, human

Most recents protocols related to «Calpeptin»

1
Calpeptin effects on fasting rats
Four male rats were randomly divided into two groups.
1) Calpeptin-administration fasting group (n = 2): Calpeptin (Cat. #14593, Cayman Chemical, Ann Arbor, MI) was dissolved in DMSO (20 mg/mL) and intraperitoneally administered at the beginning of the fasting period (0.05 mL), and then again at 24 hr after commencing fasting (0.05 mL).
2) Vehicle-administration fasting group (n = 2): rats were treated as in 1), except that the equivalent amount of vehicle (DMSO) was administered instead of the inhibitor.
Fujita Y., Taniguchi A., Yamamoto H., Obinata H., Kogo H., Iizuka-Kogo A., Ikezawa M., Tajika Y., Yokoo S, & Matsuzaki T. (2024). Aquaporin-5 Protein Is Selectively Reduced in Rat Parotid Glands under Conditions of Fasting or a Liquid Diet. Acta Histochemica et Cytochemica, 57(1), 25-33.
Publication 2024
2
ATR, ATM, Caspase, and Calpain Inhibitors
WERI-Rb1 cells were treated with ATR inhibitor VE822 (1 μM), ATM serine/threonine kinase (ATM) inhibitor KU55933 (1 μM), Caspase inhibitor Z-VAD-FMK (10 μM), or calpain inhibitor Calpeptin (10 μM), each for 1 h prior to transfection or the UV irradiation. These doses were selected based on dose-response experiments, which demonstrated that at these concentarations, the kinases were effectively inhibited without initiating an apoptotic response.
Liu J., Tan X., Li L., Cao L., Zhou Y., Li H, & Peng T. (2024). Protein expression of nucleolar protein 12 in the retina and its implication in protection of retina from UV irradiation damage. Cell Death Discovery, 10, 130.
Full text: Click here
Publication 2024
3
Modulating cAMP and Rho GTPases
To elevate intracellular cAMP concentration, 5 μM Forskolin (Sigma Aldrich Chemie, #F6886), and 10 μM Rolipram (Sigma Aldrich Chemie, #R6520) were applied for 1 h. To stimulate the activation of Rho family small GTPases cells were treated with 0.25 μg/ml Rho/Rac/Cdc42 Activator I (Cytoskeleton Inc, #CN04-A) for 2 h. Triggering of RhoA was achieved with the Rho activator I, Calpeptin at 1 U/ml (#CN01-A, Cytoskeleton Inc.). Inhibition of PKA was achieved using 10 µM Dihydrochloride (H89, Santa Cruz, sc-3537).
The following antibodies were used in this study.
Moztarzadeh S., Sepic S., Hamad I., Waschke J., Radeva M.Y, & García-Ponce A. (2024). Cortactin is in a complex with VE-cadherin and is required for endothelial adherens junction stability through Rap1/Rac1 activation. Scientific Reports, 14, 1218.
Full text: Click here
Publication 2024
4
Antiviral Compound Screening Protocol
MG132 (Selleckchem, S2619), gefitinib (Selleckchem, S1025), nigericin (InvivoGen, tlrl-nig/NIG-36-01), brefeldin A (Cell Signaling Technology, 9972S), FTY720 (Santa Cruz Biotechnology, sc-202161A), IBMX, concanamycin A (Enzo Life Sciences, ALX-380-034-C025), tetrandrine (Selleckchem, S2403), U18666A (Cayman Chemical, 10009085), ETP-46464 (Selleckchem, S8050), JIB-04 (Tocris, 4972), nitazoxanide (COVID Box, MMV688991), ketoconazole (COVID Box, MMV637533), AG-1478 (Selleckchem, S2728), caffeic acid (Selleckchem, S7414), thapsigargin (Cell Signaling Technology, 1278S), staurosporine (Cell Signaling Technology, 9953S), and arbidol-HCl (Selleckchem, S2120). Calpain Inhibitor set includes ALLN, calpain inhibitor III, calpeptin, and E-64d used in the viral inhibition assays (208733-1SET, Sigma-Aldrich).
Zeng Q., Antia A., Casorla-Perez L.A., Puray-Chavez M., Kutluay S.B., Ciorba M.A, & Ding S. (2024). Calpain-2 mediates SARS-CoV-2 entry via regulating ACE2 levels. mBio, 15(3), e02287-23.
Full text: Click here
Publication 2024
5
Microcrystal Growth Protocols for SARS-CoV-2 Mpro
Mpro microcrystals were grown using seeded batch crystallization in the XBI laboratories of the European XFEL27 . Seed crystals were grown from protein purified in the presence of 0.5 mM TCEP, using a sitting drop geometry by combining 250 nL Mpro protein solution (6.25 mg/ml) and 250 nL precipitant (25% PEG1500, 0.1 M MIB buffer pH 7.5, 5% DMSO), as reported previously28 (link). A seed stock was produced by adding the resulting Mpro crystals to a reaction tube containing a glass bead (Beads-for-Seeds, Jena Bioscience) and vortexing periodically for 5 s with subsequent incubation at room temperature. For the microcrystal batch crystallization, 250 μL glass seed beads were added to a 1.5 mL reaction tube, which was then filled with 900 μL precipitant solution (25% PEG1500, 0.1 M MIB buffer pH 7.5, 5% DMSO) mixed with 100 μL seed stock and 100 μL Mpro protein solution (35 mg/ml). Subsequently, crystals were grown in a shaker at 18 °C at 900 rpm overnight. The reduced and oxidized forms were crystallized separately. Resulting crystals were thin plates with a size ranging from 3–15 μm. Crystal concentration was adjusted by allowing the crystals to settle overnight and removing supernatant accordingly. Final crystal slurry was filtered through a 30 μm mesh gravity filter (Sysmex CellTrics) before injection.
Protein crystals for single-crystal rotation experiments were produced as previously reported28 (link), using orthorhombic seeds and reduced protein at 6.25 mg/mL. For the ligand free and S-calpeptin containing crystallization experiments, the same reduced protein batch was used. The S-calpeptin compound was dried in the well prior to crystallization mixture addition, yielding a maximum concentration of 5 mM.
Reinke P.Y., Schubert R., Oberthür D., Galchenkova M., Rahmani Mashhour A., Günther S., Chretien A., Round A., Seychell B.C., Norton-Baker B., Kim C., Schmidt C., Koua F.H., Tolstikova A., Ewert W., Peña Murillo G.E., Mills G., Kirkwood H., Brognaro H., Han H., Koliyadu J., Schulz J., Bielecki J., Lieske J., Maracke J., Knoska J., Lorenzen K., Brings L., Sikorski M., Kloos M., Vakili M., Vagovic P., Middendorf P., de Wijn R., Bean R., Letrun R., Han S., Falke S., Geng T., Sato T., Srinivasan V., Kim Y., Yefanov O.M., Gelisio L., Beck T., Doré A.S., Mancuso A.P., Betzel C., Bajt S., Redecke L., Chapman H.N., Meents A., Turk D., Hinrichs W, & Lane T.J. (2024). SARS-CoV-2 Mpro responds to oxidation by forming disulfide and NOS/SONOS bonds. Nature Communications, 15, 3827.
Full text: Click here
Publication 2024

Top products related to «Calpeptin»

1
Calpeptin by Merck Group
Sourced in United States, Germany
Calpeptin is a laboratory reagent produced by Merck Group. It functions as a potent and selective inhibitor of calpains, a family of calcium-dependent cysteine proteases. Calpains play a role in various cellular processes. Calpeptin is commonly used in research applications to study the involvement of calpains in biological systems.
2
Mg132 by Merck Group
Sourced in United States, Germany, China, United Kingdom, Macao, Sao Tome and Principe, France, Canada, Italy, Switzerland, Morocco, Belgium, Japan, Sweden, Australia, Austria, Israel, Spain
MG132 is a proteasome inhibitor, a type of laboratory reagent used in research applications. It functions by blocking the activity of the proteasome, a complex of enzymes responsible for the degradation of proteins within cells. MG132 is commonly used in cell biology and biochemistry studies to investigate the role of the proteasome in various cellular processes.
3
Calpeptin by Bio-Techne
Sourced in United States
Calpeptin is a laboratory reagent produced by Bio-Techne. It functions as a selective and potent inhibitor of the calcium-activated protease calpain.
4
Calpeptin by Selleck Chemicals
Sourced in United States, China
Calpeptin is a laboratory reagent that functions as a selective inhibitor of the calcium-activated protease calpain. It is commonly used in research applications to study the role of calpain in various cellular processes.
5
Fetal bovine serum (fbs) by Thermo Fisher Scientific
Sourced in United States, China, United Kingdom, Germany, Australia, Japan, Canada, Italy, France, Switzerland, New Zealand, Brazil, Belgium, India, Spain, Israel, Austria, Poland, Ireland, Sweden, Macao, Netherlands, Denmark, Cameroon, Singapore, Portugal, Argentina, Holy See (Vatican City State), Morocco, Uruguay, Mexico, Thailand, Sao Tome and Principe, Hungary, Panama, Hong Kong, Norway, United Arab Emirates, Czechia, Russian Federation, Chile, Moldova, Republic of, Gabon, Palestine, State of, Saudi Arabia, Senegal
Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
6
Cycloheximide (chx) by Merck Group
Sourced in United States, Germany, United Kingdom, China, Italy, Sao Tome and Principe, France, Macao, Japan, Spain, Canada, Switzerland, India, Israel, Brazil, Poland, Portugal, Australia, Morocco, Sweden, Austria, Senegal, Belgium
Cycloheximide is a laboratory reagent commonly used as a protein synthesis inhibitor. It functions by blocking translational elongation in eukaryotic cells, thereby inhibiting the production of new proteins. This compound is often utilized in research applications to study cellular processes and mechanisms related to protein synthesis.
7
Calpeptin by Cayman Chemical
Sourced in United States
Calpeptin is a laboratory reagent provided by Cayman Chemical. It functions as an inhibitor of calpain, a calcium-activated cysteine protease enzyme.
8
Dimethyl sulfoxide (dmso) by Merck Group
Sourced in United States, Germany, United Kingdom, China, Italy, Sao Tome and Principe, France, Macao, India, Canada, Switzerland, Japan, Australia, Spain, Poland, Belgium, Brazil, Czechia, Portugal, Austria, Denmark, Israel, Sweden, Ireland, Hungary, Mexico, Netherlands, Singapore, Indonesia, Slovakia, Cameroon, Norway, Thailand, Chile, Finland, Malaysia, Latvia, New Zealand, Hong Kong, Pakistan, Uruguay, Bangladesh
DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.
9
Calpeptin by Cytoskeleton
Sourced in United States
Calpeptin is a potent and selective inhibitor of the calcium-activated cysteine protease calpain. It is a widely used research tool for studying the role of calpain in various cellular processes.
10
Y 27632 by Merck Group
Sourced in United States, Germany, United Kingdom, Japan, Canada, Australia, Switzerland, Israel, China, Morocco
Y-27632 is a selective and potent Rho-associated protein kinase (ROCK) inhibitor. It functions by inhibiting the activity of ROCK, a key enzyme involved in various cellular processes.

More about "Calpeptin"

Calpeptin is a non-peptide, cell-permeable calpain inhibitor that has been extensively utilized in biological research to investigate the functions of calpain proteases.
Calpain is a calcium-dependent cysteine protease involved in various cellular processes like cytoskeletal remodeling, signal transduction, and apoptosis.
Calpeptin selectively and reversibly inhibits calpain activity, making it a valuable tool for understanding the involvment of calpain in physiological and pathological conditions, such as neurodegeneration, cancer, and inflammation.
Researchers can leverage PubCompare.ai's AI-driven platform to easily identify and compare the most relevant Calpeptin research protocols from the literature, preprints, and patents, optimizing their investigations into this important protease inhibitor.
The platform's intelligent comparisons help users locate the best Calpeptin protocols and products, streamlining the research process.
In addition to Calpeptin, other related compounds like MG132 (a proteasome inhibitor), FBS (fetal bovine serum), Cycloheximide (a protein synthesis inhibitor), DMSO (a solvent), and Y-27632 (a Rho-associated protein kinase inhibitor) may be of interest to researchers studying calpain and its role in cellular processes.
PubCompare.ai's cutting-edge technology enables seamless research by providing easy access to a wide range of relevant protocols and products.