A stock solution of BIBR (Selleck Chemicals LLC, Houston, TX, USA) at a concentration of 50 mM was prepared by dissolving the compound in sterile dimethyl sulfoxide (DMSO) and stored in small aliquots at −80 °C until use. Ammonium pyrrolidine dithiocarbamate (PDTC) (P8765; Sigma-Aldrich, Saint Louis, MO, USA) was prepared by resuspending the compound in sterile water at a concentration of 10 mM divided into aliquots and stored at −20 °C until use. Fludarabine (FLU, F9813; Sigma-Aldrich) was prepared by resuspending the compound in DMSO at a concentration of 10 mM. Cyclophosphamide (CY, 0768; Sigma-Aldrich) was prepared by dissolving the compound in sterile water at a concentration of 143.3 mM.
Ammonium pyrrolidinedithiocarbamate pdtc
Manufactured by Merck Group
Sourced in United States
Ammonium pyrrolidinedithiocarbamate (PDTC) is a chemical compound that can be used as a laboratory reagent. It functions as a metal chelating agent, capable of forming complexes with various metal ions. The core function of PDTC is to facilitate the extraction, separation, and analysis of these metal species in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (9 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Penicillin and streptomycin, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Anti nf κb p65, by Santa Cruz Biotechnology (1 mentions)
Anti p50, by Santa Cruz Biotechnology (1 mentions)
Anti cd68, by Agilent Technologies (1 mentions)
Rabbit anti sortilin, by Abcam (1 mentions)
Anti iκbα, by Santa Cruz Biotechnology (1 mentions)
Anti bcl 2, by Santa Cruz Biotechnology (1 mentions)
Oligofectamine, by Thermo Fisher Scientific (1 mentions)
Cell culture reagents, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Bcl 2 15071, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3 9664, by Cell Signaling Technology (1 mentions)
Cleaved caspase 9 9509, by Cell Signaling Technology (1 mentions)
Sirtuin antibody sampler kit, by Cell Signaling Technology (1 mentions)
18 protocols using ammonium pyrrolidinedithiocarbamate pdtc
1
Preparation of Compounds for Cell Assays
2
Comprehensive Cell Culture Reagents
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Cell culture reagents were obtained from Invitrogen (San Diego, CA, USA), unless otherwise specified. Fetal bovine serum (FBS) was from Biological Industries (Haemek, Israel) and oligofectamine and G418 from Invitrogen. NGF and BDNF from Peprothec (Rocky Hill, NJ). ProNGF from Scil Proteins GmbH (Halle, Germany) and ammonium pyrrolidinedithiocarbamate (PDTC) from Sigma-Aldrich (St. Louis, MO USA). Antibodies used in this study were the following: mouse anti-α-smooth muscle actin (α-SMA, 1∶500; Dako, Dakopatts, Denmark), anti-smooth muscle myosin (1∶400; NeoMarkers, Fremont, CA USA), anti-CD68 (Dako, 1∶250), anti-α-tubulin and anti-Chromosome Region Maintenance 1 (CRM1, 1∶1000; Sigma-Aldrich), rabbit anti-sortilin (1∶200, Abcam, Cambridge, UK; Chemicon Intern, Temecula, CA USA), anti-proNGF (Sigma-Aldrich), anti-bax protein (1∶200), anti-NF-κB p65 (1∶200), anti-p50 (1∶100), anti-IκB-α (1∶50), goat anti-p75NTR (1∶200), anti-bcl-2 (1∶100, Santa Cruz Biotechnology, CA, USA) and anti-hypoxanthine-guanine phosphoribosyltransferase (HPRT; Abcam). Fluorocrome conjugated secondary antibodies were purchased from Jackson (Suffork, UK) and Invitrogen. Horseradish peroxidase-conjugated secondary antibodies were from Nordic (Tilburg, The Netherlands).
3
Apoptosis Signaling Pathway Analysis
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The following antibodies and reagents were used: Sirtuin Antibody Sampler Kit #9787T, NF‐κB p65 Antibody Sampler Kit #4767, Death Receptor Antibody Sampler Kit #8356, antibodies against cleaved PARP #5626, cleaved caspase‐3 #9664, cleaved caspase‐8 #8592, cleaved caspase‐9 #9509, Bak #6947, Bax #5023, Bcl‐2 #15071, and Bcl‐X #2764 (Cell Signaling Technology, Danvers, MA, USA); antibodies against HDAC3 #sc‐130319 and β‐actin #sc‐58673 (Santa Cruz Biotechnology, Dallas, TX, USA); nicotinamide (NAM); and ammonium pyrrolidine dithiocarbamate (PDTC) (Sigma‐Aldrich, St Louis, MO, USA).
4
Intrathecal Delivery of TLR4 Antagonist and NF-κB Inhibitor in Rats
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The TLR4-specific antagonist TAK-242 (Millipore, 508336) was dissolved in sterile saline containing 10% DMSO. Ammonium pyrrolidinedithiocarbamate (PDTC)
, a specific inhibitor of NF-κB activation, was purchased form Sigma (St. Louis, MO) and freshly dissolved daily in normal sterile saline. Drugs were delivered by intrathecal (i.t.) or subcutaneous (s.c.) plantar injection to rats. The intrathecal catheterization was performed as previously described.24 (link),25 (link) In brief, a polyethylene-10 (outside diameter, 0.61 mm; inside diameter, 0.28 mm) catheter was inserted into the rat’s subarachnoid space through the L5–L6 intervertebral space, and the tip of the catheter was located at the L5 spinal segmental level. The doses of TAK-242 (5, 20 µg/10 µl)26 ,27 (link) and PDTC (0.2, 0.5 µg/10 µl)28 (link),29 (link) used in i.t. injection were based on the previous studies. The doses of s.c. injection of TAK-242 (20 µg/20 µl) or PDTC (0.5 µg/20 µl) were based on the doses of i.t. injection.
, a specific inhibitor of NF-κB activation, was purchased form Sigma (St. Louis, MO) and freshly dissolved daily in normal sterile saline. Drugs were delivered by intrathecal (i.t.) or subcutaneous (s.c.) plantar injection to rats. The intrathecal catheterization was performed as previously described.24 (link),25 (link) In brief, a polyethylene-10 (outside diameter, 0.61 mm; inside diameter, 0.28 mm) catheter was inserted into the rat’s subarachnoid space through the L5–L6 intervertebral space, and the tip of the catheter was located at the L5 spinal segmental level. The doses of TAK-242 (5, 20 µg/10 µl)26 ,27 (link) and PDTC (0.2, 0.5 µg/10 µl)28 (link),29 (link) used in i.t. injection were based on the previous studies. The doses of s.c. injection of TAK-242 (20 µg/20 µl) or PDTC (0.5 µg/20 µl) were based on the doses of i.t. injection.
5
Inactivated GAS Immune Response Study
6
HK2 Cell Line Culture and Treatments
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The HK2 cell line (American Type Culture Collection, ATCC-CRL2190) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 mg/mL streptomycin at 37 °C in a humidified 5% CO2 atmosphere. Ammonium pyrrolidine dithiocarbamate (PDTC) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). AAI (Subtype I of AA) was purchased from Cayman Chemical Company (Ann Arbor, MI, USA). Masson’s trichrome stain kit was purchased from ScyTek Laboratories (Logan, UT, USA). The metallothionein (MT) antibody was purchased from Invitrogen Corporation (Camarillo, CA, USA). DMEM, FBS, MEM non-essential amino acids (NEAA), penicillin, and streptomycin for cell culture were obtained from Invitrogen-Gibco (Grand Island, NY, USA).
7
Mesangial Cell Response to High Glucose
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Mouse glomerular mesangial cells SV40-MES-13 were purchased from ATCC and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Gibco) in low glucose (5.5 mM of glucose) supplemented with 10% fetal bovine serum (FBS, Gibco) with 1% antibiotics (100 IU/ml penicillin-streptomycin), and then grown at 37°C with an atmosphere of 5% CO2. Next, cells were treated with SA (10, 20, and 30 μM) dissolved in 0.1% dimethyl sulfoxide for 1 h before high glucose (30 mM, Sigma) treatment. After 24 h of culture, the supernatant and cells were collected for further analysis. To further explore the mechanism, mesangial cells were pretreated with ammonium pyrrolidine dithiocarbamate (PDTC, 50 μM, Sigma), a NF-κB inhibitor, along with SA prior to stimulating with HG for 24 h.
8
Liver Cancer Cell Line Culture
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The human hepatoma cell lines (SMMC-7721, Bel-7402, Hep3B and MHCC-97H) and non-malignant liver cells (LO2) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and cultured in High glucose Dulbecco’s Modified Eagle Medium (DMEM) (HyClone, Shanghai, China) supplemented with 10% fetal bovine serum (FBS) (Invitrogen, Carlsbad, CA, USA), with 100 U/ml of penicillin G and 100 μg/ml of streptomycin at 37°C in a water-saturated atmosphere of 5% CO2. For studies assessing the effect of NF-κB inhibition, cells were treated the NF-κB inhibitor ammonium pyrrolidinedithiocarbamate (PDTC) (Sigma-Aldrich, Poole, UK) at a final concentration of 20 μM.
9
Inhibition of NF-κB and Ghrelin Signaling in Paclitaxel-Treated Rats
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Paclitaxel (LKT Laboratories inc, St. Paul, MN, USA) was dissolved in 10% dimethyl sulfoxide (10%DMSO). Ammonium pyrrolidine dithiocarbamate (PDTC; Sigma-Aldrich, St Louis, MO, USA), an NFκB inhibitor, and [D-Lys3]-GHRP-6 (Tocris, Bristol, United Kingdom), a ghrelin receptor antagonist, were dissolved in saline. RKT (Tsumura, Tokyo, Japan) was dissolved in distilled water (DW).
Paclitaxel was injected intraperitoneally (i.p.) at a dose of 2 or 5 mg/kg. PDTC was injected intrathecally (i.t.) at a dose of 100 ng/5μL. [D-Lys3]-GHRP-6 was injected i.p. at a dose of 10 mg/kg. RKT was administered orally (p.o.) at a dose of 0.1, 0.3, or 1 g/kg. Control animals were injected with the respective vehicle for each drug.
Paclitaxel was injected intraperitoneally (i.p.) at a dose of 2 or 5 mg/kg. PDTC was injected intrathecally (i.t.) at a dose of 100 ng/5μL. [D-Lys3]-GHRP-6 was injected i.p. at a dose of 10 mg/kg. RKT was administered orally (p.o.) at a dose of 0.1, 0.3, or 1 g/kg. Control animals were injected with the respective vehicle for each drug.
10
Oxaliplatin-induced Thermal Hyperalgesia Model
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Oxaliplatin was purchased from Sigma (USA) and dissolved in 5% glucose/H2O as a stock solution of 1 mg/ml, as reported in a previous study.8 (link) Rats were intraperitoneally administered Oxaliplatin at 4 mg/kg once per day for five consecutive days to induce thermal hyperalgesia. The control rats were intraperitoneally injected with an equivalent volume of 5% glucose/H2O. Intrathecal injection of isotype IgG (10 μg in 10 μl, R&D system, USA), a neutralizing antibody against CX3CL1 (10 μg in 10 μl, Torrey Pines Bio Labs, USA), ammonium pyrrolidine dithiocarbamate (PDTC; 200 ng in 10 μl, Sigma, USA), scramble siRNA (50 μg in 15 μl, Ribobio, China), or NF-κB p65 small interfering RNA (siRNA; 50 μg in 15 μl, Ribobio, China) was performed 30 min before Oxaliplatin administration.
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