Pyrrolidine dithiocarbamate

Manufactured by Beyotime

Sourced in China

Pyrrolidine dithiocarbamate is a chemical compound used in laboratory settings. It functions as a metal chelator, capable of forming stable complexes with various metal ions.

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Lab products found in correlation

4 protocols using pyrrolidine dithiocarbamate

Quercetin Inhibits Inflammatory Signaling

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Quercetin (purity: ≥98%) was purchased from Sigma Chemical Co. (St. Louis, MO), and it was dissolved in dimethyl sulfoxide (DMSO) for the next experiments. Fetal bovine serum (FBS) and Dulbecco modified Eagle medium (DMEM) were obtained from GIBCO BRL Co. (Gaithersburg, MD, USA). Rabbit antihuman TLR4 antibody was obtained from AMS Biotechnology (Abington, UK), whereas pyrrolidine dithiocarbamate (PDTC; an NF-κB inhibitor) was purchased from Beyotime Institute of Biotechnology (Nantong, China). The antibodies to TLR4, NF-κB p65, mitochondrial membrane potential-2 (MMP-2), and MMP-9 were obtained from Cell Signaling Technology (Beverly, MA, USA). The enzyme-linked immunosorbent assay (ELISA) kits for tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (Cox-2), and interleukin-6 (IL-6) were purchased from KeyGEN Biotech. Co., LTD. (Nanjing, China). Growth factor-reduced Matrigel was purchased from Becton Dickinson (BD Biosciences, San Jose, USA). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from Sigma Chemical Co.(St. Louis, MO, USA). All other materials were from commercial sources.

Han M., Song Y, & Zhang X. (2016). Quercetin Suppresses the Migration and Invasion in Human Colon Cancer Caco-2 Cells Through Regulating Toll-like Receptor 4/Nuclear Factor-kappa B Pathway. Pharmacognosy Magazine, 12(Suppl 2), S237-S244.

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Molecular Mechanisms of Neuroinflammation

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ATX (Santa Cruz Biotechnology, Santa Cruz, CA, USA, 98% pure) was dissolved in dimethyl sulfoxide (DMSO); the DMSO content in every group was 0.1%. Pyrrolidine dithiocarbamate (PDTC), a NF-кB inhibitor, was purchased from Beyotime (Jiangsu, China) and used at a concentration of 10 μmol/L. Bumetanide, a NKCC1-specific inhibitor, was purchased from Sigma-Aldrich (St. Louis, MO, USA) and used at a concentration of 50 μmol/L [22 (link)]. Anti-NKCC1, anti-glial fibrillary acidic protein (GFAP) and anti-NF-кB/p65 antibodies were purchased from Millipore (Billerica, MA, USA) and Santa Cruz Biotechnology, respectively. Anti-pro caspase 3 antibody was purchased from Abcam (Cambridge, MA, USA). Antibodies against Bcl-2, Bax, cleaved caspase-3, IL-1β, IL-6, TNF-α, β-actin, β-tubulin, and Histone H3 were purchased from Cell Signaling Technology (Danvers, MA, USA).

Zhang M., Cui Z., Cui H., Wang Y, & Zhong C. (2017). Astaxanthin protects astrocytes against trauma-induced apoptosis through inhibition of NKCC1 expression via the NF-κB signaling pathway. BMC Neuroscience, 18, 42.

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Inhibition of NF-κB in Stretched Myoblasts

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Two selective and irreversible NF-κB inhibitors: BAY11-7082 (BAY; Beyotime Institute of Biotechnology, Jiangsu, China) and pyrrolidine dithiocarbamate (PDTC; Beyotime Institute of Biotechnology) were used in the present study. In order to determine the effects of NF-κB inhibition on the stretched myoblasts, C2C12 cells that underwent 0.125 Hz cyclic strain (10% deformation) for 2 h per day for 4 consecutive days were simultaneously treated with 2.5 µM BAY, 5 µM BAY or 10 µM PDTC. The effects of NF-κB inhibition on stretched C2C12 myoblasts were subsequently detected according to the aforementioned CCK-8 method and FCM analysis at corresponding time points.

Hua W., Zhang M., Wang Y., Yu L., Zhao T., Qiu X, & Wang L. (2016). Mechanical stretch regulates microRNA expression profile via NF-κB activation in C2C12 myoblasts. Molecular Medicine Reports, 14(6), 5084-5092.

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Isolation and culture of primary podocytes

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Bilateral kidneys were placed in DMEM/F12 culture medium, the envelope was removed, the kidney cortex was separated on ice, and the tissue was collected on a 200 mesh sieve, centrifuged at 1,500 r/min for 5 min, resuspended in 2 mL of complete medium and then inoculated into culture flasks, incubated at 37°C, 5% CO 2 incubator for 5 h [18] . Cell morphology was observed microscopically. Podocytes were identified by immunofluorescent staining for podocalyxin and WT-1. Podocytes were inoculated in culture dishes for 24 h in serum-free culture. Si-PL2R (AAG GCA GTT CTG GAT TGG ATT TAA T) was transfected to the primary podocytes for 24-48 h. Recombinant PLA2G12B protein at the concentration of 100 ng/mL was applied and incubated with podocytes. Subsequently, 10 μmol/L NF-κB inhibitor pyrrolidinedithiocarbamate (Beyotime, S1808) was added. Each group of cells was collected.

Fu L., Ping J., Guo F., Song J., Luo M, & Chen L. (2023). PLA2G12B Mediates Arachidonic Acid Metabolism through Activation of the NF-κB Pathway to Promote Membrane Nephropathy. Kidney & blood pressure research, 48(1).

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