Calpeptin

Manufactured by MedChemExpress

Sourced in China, United States

Calpeptin is a laboratory reagent that functions as a selective and potent inhibitor of the calcium-activated neutral protease calpain. It binds to and inhibits the activity of calpain, a family of enzymes involved in various cellular processes.

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Lab products found in correlation

5 protocols using calpeptin

Cysteine Protease Inhibitor Screening

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Boceprevir, Telaprevir, Narlaprevir, Grazoprevir, Asunaprevir, Simeprevir, Leupeptin, VBY-825, Balicatib, CA-074, Calpeptin, E-64, E-64c, JPM-OEt, LY-3000328, Odanacatib, CPI (Cysteine Protease Inhibitor), Aloxistatin, Cinanserin, MDL-28170, MG-101, ONO-5334, (±)Alliin, and Z-LVG-CHN2 were purchased from MedChemExpress, LLC. (www.medchemexpress.com). DiscoveryProbe™ FDA-approved Drug Library (23 plates of 96-well, 1971 compounds) were purchased from APExBIO (https://www.apexbt.com).

Andi B., Kumaran D., Kreitler D.F., Soares A.S., Keereetaweep J., Jakoncic J., Lazo E.O., Shi W., Fuchs M.R., Sweet R.M., Shanklin J., Adams P.D., Schmidt J.G., Head M.S, & McSweeney S. (2022). Hepatitis C virus NS3/4A inhibitors and other drug-like compounds as covalent binders of SARS-CoV-2 main protease. Scientific Reports, 12, 12197.

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Screening Potential SARS-CoV-2 Inhibitors

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We purchased
HEK293T/17 cells from ATCC; DMEM with high glucose
with GlutaMAX supplement, fetal bovine serum, 0.25% trypsin-EDTA,
phenol red, puromycin, Lipofectamine 3000, and dimethyl sulfoxide
from Thermo Fisher Scientific; linear polyethylenimine MW 25000 from
Polysciences; RealTime-Glo annexin V apoptosis and a necrosis assay
kit from Promega; an EndoFree plasmid DNA midi kit from Omega Biotek;
antimycin a from Sigma-Aldrich; GC376 from Selleck Chem; boceprevir,
calpeptin, MG-132, telaprevir, and carmofur from MedChemExpress; ebselen
from TCI; calpain inhibitors II and XII from Santa Cruz Biotechnology;
MG-115 From Abcam; tideglusib, disulfiram, and PX-12 from Cayman Chemical;
chloroquine diphosphate from Alfa Aesar; hydroxychloroquine sulfate
from Acros Organics; and a fluorogenic M^Pro substrate DABCYL-Lys-Thr-Ser-Ala-Val-Leu-Gln-Ser-Gly-Phe-Arg-Lys-Met-Glu-EDANS
termed as Sub3 from Bachem. K777 was a gift from Prof. Thomas Meek
at Texas A&M University. The syntheses of MPI1–9 and 11a
were shown in a previous publication.^{15 (link)}

Cao W., Cho C.C., Geng Z.Z., Shaabani N., Ma X.R., Vatansever E.C., Alugubelli Y.R., Ma Y., Chaki S.P., Ellenburg W.H., Yang K.S., Qiao Y., Allen R., Neuman B.W., Ji H., Xu S, & Liu W.R. (2022). Evaluation of SARS-CoV-2 Main Protease Inhibitors Using a Novel Cell-Based Assay. ACS Central Science, 8(2), 192-204.

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RPMI-2650 Cell Line Exposure to Cigarette Smoke and Inflammatory Stimuli

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The human nasal epithelial cell line, RPMI-2650, was purchased from the Cell Bank of the Chinese Academy of Sciences. The cells were cultured in RPMI 1640 medium (Hyclone, Logan, Utah, USA) supplemented with 10% foetal bovine serum (Gibco, CA, USA) and 1% penicillin-streptomycin solution (Biosharp, China), and placed in a 5% CO₂ humidified environment at 37 °C. The RPMI-2650 cells were inoculated in six-well plates at a density of 1.0 × 10⁵ cells per well. Either CSE (18.02%) or control medium was added to the cell cultures, which were grown to 60–70% confluence. After 24 h, the cell culture medium was changed, and the cells were stimulated overnight with TNF-α (10 ng/mL; PeproTech, New Jersey, USA) [17 ]. Next, the cells were divided into four groups: 1) control medium + TNF-α stimulation; 2) CSE medium + TNF-α stimulation; 3) CSE medium + TNF-α stimulation + Dexamethasone; and 4) CSE medium + TNF-α stimulation + calpeptin. Dexamethasone (2.51 × 10⁻⁶ M) [17 ] and calpeptin (50 nmol, MedChemExpress, China) [11 (link)] were each administered for 1 h.

Zhu W.X., Xi Y., Li F., Jiao W.E., Li Z.J., Chen S.M., Kong Y.G., Xu Y., Deng Y.Q., Zuo J.J, & Tao Z.Z. (2023). Calpeptin may reverse glucocorticoid-resistance of allergic rhinitis associated with cigarette smoke exposure by down-regulating interferon regulatory factor 1. Heliyon, 9(7), e17316.

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Calpain Activity Quantification in PDAC Cells

Cited 1 time

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Calpain activity was assessed using the Calpain-Glo^™ assay kit obtained from Promega (Madison, WI, G8502) in accordance with the manufacturer’s instructions and based on previous studies.^{64 (link),65 (link)} PDAC cells (AsPC-1 1x10⁴ cells; BxPC-3 0.6x10⁴ cells; Panc 10.05 2x10⁴ cells) were seeded in a 96-well white plate (Thermo Fisher Scientific) with regular growth media and incubated for 24 hours. The Calpain-Glo reagent^™ and Suc-LLVY-Glo^™ substrate were mixed and incubated without Ca²⁺. Subsequently, the cell culture medium was aspirated, and 90 μl of the mixture was added to each well, followed by a 20-minute incubation at 37°C. For cell lysis, 10 μl of lysis buffer containing 9% Triton-X-100 and 1 mM of the calpain inhibitor Calpeptin (MedChemExpress LLC, Monmouth Junction, NJ) was introduced to the wells to prevent further substrate cleavage by calpain. Luminescence, indicative of cellular calpain activity, was measured using the EnSpire^™ multimode plate reader (PerkinElmer, Waltham, MA).

Ohara Y., Tang W., Liu H., Yang S., Dorsey T.H., Cawley H., Moreno P., Chari R., Guest M.R., Azizian A., Gaedcke J., Ghadimi M., Hanna N., Ambs S, & Hussain S.P. (2023). SERPINB3-MYC axis induces the basal-like/squamous subtype and enhances disease progression in pancreatic cancer. Cell reports, 42(12), 113434.

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Oxidative Stress Modulation Protocol

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FW, containing 20% Cl₂ and 80% hypochlorous acid (pH, 2.2-2.7) was obtained using the Oxilyzer (Miura Denshi, Akita, Japan). Cycloheximide (CHX) was purchased from Sigma-Aldrich (Tokyo, Japan). The calpain inhibitor calpeptin was purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Takemoto T., Kaetsu R., Hanayama M., Ishiyama Y., Sadamura M., Nishio K., Tsunoda M., Asano M, & Motoyoshi M. (2021). Acid-electrolyzed functional water-induces Interleukin-1α release from Intracellular Storage Sites in Oral Squamous Cell Carcinoma. International Journal of Medical Sciences, 18(8), 1746-1752.

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