Copper 2 sulfate solution

Manufactured by Merck Group

Sourced in United States

Copper (II) sulfate solution is a chemical compound consisting of copper and sulfate ions dissolved in water. It is a blue-colored liquid commonly used in various laboratory applications.

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Copper sulfate (142 citations) Copper(II) sulfate (61 citations) Copper (II) sulfate pentahydrate 4% solution (3 citations)

9 protocols using copper 2 sulfate solution

Protein Extraction and Analysis from Jurkat T and Ramos B Cells

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After treatment with fludioxonil in 10⁻⁷ M–10⁻⁵ M cells, proteins from the Jurkat T cells and Ramos B cells were extracted by radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 1% NP-40, 0.5% deoxycholic acid, and 0.1% SDS). The protein concentrations were determined by a solution of bicinchoninic acid (BCA; Sigma-Aldrich Corp.) which was mixed with copper II sulfate solution (Sigma-Aldrich Corp.) at a ratio of 50:1. The total cell proteins (30 µg) were separated in a 15% SDS-PAGE gel and transferred to a polyvinylidene fluoride (PVDF) membrane (BioRad Laboratories, Hercules, CA, USA). The membrane was immersed in primary antibodies in tris-buffered saline (TBS) containing 5% bovine serum albumin (BSA; RMBIO Corp., Missoula, MT, USA), as shown in Table 1, and incubated overnight at 4 °C. The membrane was incubated with horseradish peroxidase (HRP)-conjugated secondary antibody (goat anti-mouse IgG or goat anti-rabbit IgG; BioRad Laboratories) for 2 h. proteins were detected by LuminoGraph II (ATTO, Tokyo, Japan) using EZwestLumi plus (ATTO).

Lee G.H., Hwang K.A, & Choi K.C. (2019). Effects of Fludioxonil on the Cell Growth and Apoptosis in T and B Lymphocytes. Biomolecules, 9(9), 500.

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Immunohistochemical Analysis of Serotonin System

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The following reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA); glycerol, 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI), aqueous mounting buffer, sodium hydroxide, tetramethylethylenediamine (TEMED), copper(II) sulfate solution, LPS from Escherichia coli O111:B4, sucrose, Tween 20, and bovine serum albumin (BSA). Other reagents were obtained from the following manufacturers: Triton X-100, paraformaldehyde powder, acetyl alcohol, hydrogen peroxide (H₂O₂), methylene alcohol, butanol, isopentene, sulfuric acid, phosphate, and ethylene glycol (Duksan Science, Seoul, Korea; Daejung Chemicals & Metals Co., Siheung, Korea; Junsei Chemical Co., Ltd., Tokyo, Japan), optimal cutting temperature (OCT) compound, protease inhibitor, normal goat serum, 5-hydroxytryptamin (5-HT), tryptophan hydroxylase 2 (TPH2), glial fibrillary acidic protein (GFAP), 5-HT transporter, 5HT_1A receptor, β-actin, and fluorescence- and horseradish peroxidase (HRP)-conjugated secondary antibodies (Abcam, Cambridge, MA, USA; Thermo-Fisher Scientific, Allentown, PA, USA; Santa Cruz Biotechnology, Dallas, TX, USA); and AMRESCO and RNAlater (Ambion, Austin, TX, USA).

Lee J.S., Jeon Y.J., Park S.Y, & Son C.G. (2020). An Adrenalectomy Mouse Model Reflecting Clinical Features for Chronic Fatigue Syndrome. Biomolecules, 10(1), 71.

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Western Blot Analysis of Protein Expression

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Proteins were isolated using the NucleoSpin^® RNA/Protein kit (MACHEREY-NAGEL). Protein contents were measured using Bicinchoninic acid Solution (Sigma, USA) and Copper (II) sulfate solution (Sigma, USA). Protein were separated on 12% or 8% SDS-PAGE gel. Western blotting of SDS-PAGE gels was performed using standard methodology. Primary antibodies were diluted at 1:1000 in 1% skim milk in PBST. Primary antibodies used in this study were LY6K (Santa Cruz Biotechnology, USA), ERα (Santa Cruz Biotechnology, USA), Argonate2 (abcam^®, UK) and β-actin (Bethyl Laboratories, US). β-actin was used as a loading control. Immunoreactive proteins were detected by horseradish peroxidase-conjugated secondary antibodies and the enhanced chemiluminescence reagent, EzWestLumi plus (ATTO, JAPAN).

Kim Y.S., Park S.J., Lee Y.S., Kong H.K, & Park J.H. (2016). miRNAs involved in LY6K and estrogen receptor α contribute to tamoxifen-susceptibility in breast cancer. Oncotarget, 7(27), 42261-42273.

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Mitochondrial Respiration Measurement in Hematological Cells

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OCR was assessed using a Seahorse XF96 flux analyzer (Seahorse Bioscience). KG1 (75 × 10³ cells/well) and TEX (1 × 10⁵ cells/well) cells were cultured in a 96-well XF96 culture plate (Seahorse Bioscience) coated with BD Cell-Tak (BD Biosciences) in their regular growth medium for 24 h with or without R406. An hour prior to the analysis, cells were washed with PBS and resuspended in XF base medium (pH 7.4) (Agilent Technologies) supplemented with glucose (5 mM), L-glutamine (1.6 mM), and pyruvate (1 mM). Plates were incubated for 1 h at 37 °C in a CO₂-free incubator and transferred to the XF96 analyzer. OCR was measured at baseline and after the injection of oligomycin (1 µg/ml), FCCP (1.5 µM for TEX and 1.25 µM for KG1), rotenone/antimycin A (1 µM). After measurements, cells were washed with PBS, lysed in 10 μL of 0.1% Triton/PBS solution and frozen at −80 °C overnight. Thereafter, the protein concentration in each well was measured using BCA (Copper(II) sulfate solution and Bicinchoninic Acid solution, Sigma-Aldrich). Data were normalized to protein concentration and analyzed using Wave 2.0 software (Agilent Technologies).

Polak A., Bialopiotrowicz E., Krzymieniewska B., Wozniak J., Stojak M., Cybulska M., Kaniuga E., Mikula M., Jablonska E., Gorniak P., Noyszewska-Kania M., Szydlowski M., Piechna K., Piwocka K., Bugajski L., Lech-Maranda E., Barankiewicz J., Kolkowska-Lesniak A., Patkowska E., Glodkowska-Mrowka E., Baran N, & Juszczynski P. (2020). SYK inhibition targets acute myeloid leukemia stem cells by blocking their oxidative metabolism. Cell Death & Disease, 11(11), 956.

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Cytotoxicity Assay Protocol

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DMEM, PBS (sterile), sodium pyruvate, penicillin/streptomycin, protein standard, bicinchoninic acid solution and copper (II) sulfate solution for total protein determination were purchased from Sigma-Aldrich (Taufkirchen, Germany); fetal bovine serum and glutamine were from Biochrom (Berlin, Germany). Resazurin was from Sigma Aldrich (Taufkirchen, Germany) and Alamar blue kit from BioRad (Feldkirchen, Germany) (BUF 012-B). LDH-Kit was purchased from Sigma-Aldrich (MAK066-1KT) with an additional standard from Cayman-Chemical (Michigan, USA). For the GSH enzymatic recycling method potassium dihydrogen orthophosphate, dipotassium hydrogen orthophosphate and Triton X-100 were aquired from Carl Roth (Karlsruhe, Germany). Sulfosalicylic acid was purchased from Sigma-Aldrich (Taufkirchen, Germany). DTNB, ß-NADPH, Glutathione reductase and GSH were purchased from Merck (Darmstadt, Germany). Diethiothreitol and Monobrombimane were from Sigma Aldrich (Taufkirchen, Germany).

Küstner M.J., Eckstein D., Brauer D., Mai P., Hampl J., Weise F., Schuhmann B., Hause G., Glahn F., Foth H, & Schober A. (2024). Modular air–liquid interface aerosol exposure system (MALIES) to study toxicity of nanoparticle aerosols in 3D-cultured A549 cells in vitro. Archives of Toxicology, 98(4), 1061-1080.

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Western Blotting of Vascular Smooth Muscle Cells

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The vascular smooth muscle cells were lysed with RIPA buffer (R0278, Sigma-Aldrich™). The concentrations of protein in the cell lysates were measured using Bicinchoninic Acid Kit (Cat #B9643, Sigma-Aldrich™) and copper (II) sulfate solution (C2284, Sigma-Aldrich™). For SDS PAGE, 4–15% Mini-PROTEAN® TGXTM Precast Protein Gels (456–1084, BIO-RADTM) were used. Electrophoresis was performed for a period of 90 minutes at 100 volts, or until the dye front reached the bottom end of the gel. Subsequently, the resolved proteins in the gel were transferred onto PVDF Transfer Membrane, 0.45 μm (88518, ThermoFisher™). The transferred blot was then blocked in Tris-buffered saline with Tween-20 (TBST) buffer containing 3% Bovine Serum Albumin (A7906, Sigma-Aldrich™) for 1 hour. The membrane was then probed with Primary antibody specific to CTSL (1:200, LS-C293267, LifeSpan Biosciences™) or GAPDH Antibody (1D4) (1:1000, NB300-221, Novus Biologicals™) for 16h at 4°C. After three TBST washings, the membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary antibody at a 1:5000 dilution of rabbit anti-mouse IgG Antibody [HRP] (NB7544, Novus Biologicals™) or goat anti-rabbit IgG antibody (NB7160, Novus Biologicals™) for 1 hour. PierceTM ECL Western Blotting Substrate (32106, ThermoFisher™) was used to detect protein expression on the membrane through chemiluminescence.

Gunasekar P., Satish M., Dabestani P.J., Jiang W., Boosani C.S., Radwan M., Agrawal D.K, & Asensio J.A. (2019). Modulation of Cathepsin L expression in the coronary arteries of atherosclerotic swine.. The Journal of surgical research, 243, 460-468.

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Adipocyte Differentiation and Autophagy Assay

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D-Phenylalanine (Cat# P1751), L-Leucine methyl ester hydrochloride (Cat# L1002), L-γ-glutamyl-p-nitoanilide (γ-GPNA) (Cat# G6133), L-Leucine (Cat# L8000), cycloheximide (Cat# C4859), puromycin (Cat# P9620), polybrene (Cat# H9268), doxycycline (Cat# D9891), Protein A Sepharose beads (Cat# P9424), Anti-FLAG M2 Magnetic Beads (Cat# M8823), bicinchonic acid solution (BCA) (Cat# B9643), Copper (II) sulfate solution (Cat# C2284), troglitazone (Cat# T2573), insulin (Cat# I9278), IBMX (Cat# I5879), dexamethasone (Cat# D4902), Oil Red O solution (Cat# O1391), cOmplete protease inhibitor tablets (Cat# 4693124001) and Earle’s Balanced Salt Solution (EBSS) (Cat# E2888) were obtained from Sigma-Aldrich. Fetal Bovin Serum (FBS) (Cat# 10270-106), penicillin/streptomycin (Cat#15140-122), glutamine (Cat#25030-024), accutase (Cat#A11105-01), trypsin (Cat# 15090-046), Dulbecco’s Modified Eagle Medium (DMEM) (Cat# 21969-035), LysoTracker Deep Red (cat#L12492) and 50X MEM amino acid solution (Cat# 11130-036) were purchased from Thermo Fisher Scientific. Rapamycin (Cat# R-5000) and Bafilomycin A1 (Cat# B-1080) were obtained from LC laboratories.

Beaumatin F., O’Prey J., Barthet V.J., Zunino B., Parvy J.P., Bachmann A.M., O’Prey M., Kania E., Gonzalez P.S., Macintosh R., Lao L.Y., Nixon C., Lopez J., Long J.S., Tait S.W, & Ryan K.M. (2019). mTORC1 Activation Requires DRAM-1 by Facilitating Lysosomal Amino Acid Efflux. Molecular Cell, 76(1), 163-176.e8.

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Bicinchoninic Acid Protein Assay

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Total protein was determined using the bicinchoninic acid assay (BCS - Bicinchoninic acid solution, Sigma, Saint Louis, MO, USA; Copper (II) sulfate solution, Sigma-Aldrich, Saint Louis, MO, USA). The intra-assay variability of each assay was < 5% (inter-assay CV% < 10%).

Engeland C.G., Hugo F.N., Hilgert J.B., Nascimento G.G., Junges R., Lim H.J., Marucha P.T, & Bosch J.A. (2015). The effects of psychological distress on salivary secretory immunity. Brain, behavior, and immunity, 52, 11-17.

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Carbohydrate Characterization Protocols

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d-mannose, d-galactose, d-xylose, l-rhamnose, d-glucose, d-arabinose, d-fucose, d-fructose, calcium chloride anhydrous, sodium phosphate dibasic, sodium hydroxide, barium chloride anhydrous, 1-Phenyl-3-methyl-5-pyrazolone (PMP), Folin–Denis’ reagent, gallic acid, trifluoroacetic acid (TFA), trichloroacetic acid (TCA), phenol, chloroform, sulfuric acid, nitric acid, hydrogen chloride (1N), hydrogen peroxide (30%), gelatin, copper(II) sulfate solution (4%, w/v), and bicinchoninic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bovine serum albumin standard was purchased from Thermo Fisher Scientific (Waltham, MA, USA). All analytical grade organic solvents (acetonitrile, ethanol, methanol, deionized water) were purchased from Burdick & Jackson chemicals (Muskegon, MI, USA). Ultra-pure argon (99.99%), nitrogen (99.99%), and carbon dioxide were supplied from Daechang Gas (Songha-dong, Gwangju, South Korea).

Yim S.K., Kim K., Kim I., Chun S., Oh T., Kim J.U., Kim J., Jung W., Moon H., Ku B, & Jung K. (2021). Inhibition of SARS-CoV-2 Virus Entry by the Crude Polysaccharides of Seaweeds and Abalone Viscera In Vitro. Marine Drugs, 19(4), 219.

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