The primary antibodies for HPSE (latent precursor: 65 kDa, active HPSE: 50 kDa), GAPDH, and isotype IgG were purchased from Santa Cruz (Dallas, Texas, USA); the primary antibodies for lamin B1, NF-κB p65, IκBα, and IκBα pSer32 from Cell Signaling Technology (Danvers, Massachusetts, USA); a primary antibody for heparan sulfate proteoglycan was purchased from Bioss (Woburn, Massachusetts, USA); blocking antibodies against TLR1 (GD2.F4), TLR2 (TL2.1), and TLR4 (HTA125) from eBioscience (San Diego, CA, USA); a blocking antibody against TLR6 (TLR6.127) from Abcam (Cambridge, UK); a mouse Histone H4 ELISA kit was purchased from USCN Life Science (Wuhan, China); a mouse HPSE ELISA kit was purchased from Biorbyt (Cambridge, UK). A blocking antibody against Histone H4 (anti-H4) was prepared following the previously described protocol [15 (link)]. Histone H4 was purchased from Millipore (Billerica, MA, USA); and the NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) was obtained from Abcam (Waltham, MA, USA). All other reagents were purchased from Sigma (St. Louis, MO, USA), unless stated otherwise.
Pyrrolidine dithiocarbamate
Manufactured by Abcam
Sourced in United States
Pyrrolidine dithiocarbamate is a chemical compound that is often used in laboratory research. It has the chemical formula C5H9NS2. The core function of this product is to act as a chelating agent, which means it can bind to and sequester metal ions.
Automatically generated - may contain errors
Lab products found in correlation
Isotype igg, by Santa Cruz Biotechnology (1 mentions)
Iκbα pser32, by Cell Signaling Technology (1 mentions)
Histone h4, by Merck Group (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Recombinant human il 1β, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Collagenase 2, by Merck Group (1 mentions)
Curcumin, by Merck Group (1 mentions)
Cell culture reagents, by Thermo Fisher Scientific (1 mentions)
Ab39012, by Abcam (1 mentions)
Ab7097, by Abcam (1 mentions)
Ab8245, by Abcam (1 mentions)
Ab16048, by Abcam (1 mentions)
Urb597, by Cayman Chemical (1 mentions)
No 38450, by Ugo Basile (1 mentions)
4 protocols using pyrrolidine dithiocarbamate
1
Investigating HPSE and NF-κB Signaling Pathways
2
Curcumin Modulates NF-κB Pathway in Chondrocytes
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Curcumin and collagenase II were obtained from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany) and a 10 mM stock solution was prepared in dimethyl sulfoxide, and diluted with cell culture medium immediately prior to use.
Cells were maintained in Dulbecco's modified Eagle's medium (DMEM; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco; ThermoFisher Scientific, Inc.) and 100 IU/ml penicillin, 100 mg/ml streptomycin at 37°C in a humidified atmosphere of 95% air and 5% CO2.
Recombinant human IL-1β was supplied by PeproTech, Inc. (Rocky Hill, NJ, USA). Primary antibodies against type II collagen (AB746) and MMP-13 (AB39012) were purchased from Abcam (Cambridge, UK). Primary antibodies against NF-κB inhibitor α (IκBα; 4814S), phosphorylated (p)-IκBα (9246S) and NF-κB p65/RelA (8242S) were provided by Cell Signaling Technology, Inc. (Danvers, MA, USA). The selective NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC), primary antibodies against GAPDH (AB8245) and lamin B1 (AB16048), and horseradish peroxidase (HRP)-conjugated secondary antibody (AB7097) were obtained from Abcam. Cell culture reagents were purchased from Gibco (Thermo Fisher Scientific, Inc.).
Cells were maintained in Dulbecco's modified Eagle's medium (DMEM; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco; ThermoFisher Scientific, Inc.) and 100 IU/ml penicillin, 100 mg/ml streptomycin at 37°C in a humidified atmosphere of 95% air and 5% CO2.
Recombinant human IL-1β was supplied by PeproTech, Inc. (Rocky Hill, NJ, USA). Primary antibodies against type II collagen (AB746) and MMP-13 (AB39012) were purchased from Abcam (Cambridge, UK). Primary antibodies against NF-κB inhibitor α (IκBα; 4814S), phosphorylated (p)-IκBα (9246S) and NF-κB p65/RelA (8242S) were provided by Cell Signaling Technology, Inc. (Danvers, MA, USA). The selective NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC), primary antibodies against GAPDH (AB8245) and lamin B1 (AB16048), and horseradish peroxidase (HRP)-conjugated secondary antibody (AB7097) were obtained from Abcam. Cell culture reagents were purchased from Gibco (Thermo Fisher Scientific, Inc.).
3
Regulation of VEGF and HMGB1 Release
4
Mechanical Allodynia Assessment and Pharmacological Interventions
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Mechanical allodynia was assessed by measuring mechanical withdrawal threshold values using an electronic von Frey filament (no.38450; UGO Basile, Varese, Italy). Rats were individually placed in an acrylic cage and habituated for 10 minutes. The filament was vertically applied to the planta skin of the left hind paw, and force values were measured until the animal exhibited withdrawal or licking of the hind paw. The measurements were performed seven times in 2- to 3-min intervals, and averaged data were collected, omitting the maximum and minimum values. All of the von Frey tests were assessed by a researcher who was blind to the experimental groups. For drug injections, URB597 (0.3 mg/kg, Cayman Chemical, Ann Arbor, MI, US) and PDTC (pyrrolidine dithiocarbamate, 200 mg/kg, Abcam, Cambridge, UK) were administered intraperitoneally daily for 3 days after the induction of CPIP. Additionally, hydralazine was administered at 2 mL/kg for 3 days, which constituted a dose of 25 mg/kg of hydralazine. Nocifensive behavior was tested before drug injection and at 5 days post initial injection. During the 5 days, each drug was injected on days 0–2.
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