The following reagents were purchased for the experiments of this study: Rabbit anti-mouse HO-1 antibody (Boster Biological Technology, Wuhan, China), anti-rabbit IgG (MaxVisionTM2; Fuzhou Maixin Biotechnology Development Co., Ltd., Fuzhou, China), Takara RNA polymerase chain reaction (PCR) kit version 3.0 (Avian Myeloblastosis Virus; Takara Bio, Inc., Dalian, China), hydroxyproline (HYP; Nanjing KeyGen Biotech. Co., Ltd., Nanjing, China), cobalt protoporphyrin (CoPP; Sigma-Aldrich, St. Louis, MO, USA) and octreotide (Novartis International AG, Basel, Switzerland).
Cobalt protoporphyrin copp
Manufactured by Merck Group
Sourced in China, Canada, United States
Cobalt protoporphyrin (CoPP) is a synthetic metalloporphyrin compound. It is a coordination complex consisting of a cobalt ion centered within a protoporphyrin IX ligand. CoPP is used as a laboratory reagent in biochemical and cell biology research applications.
Automatically generated - may contain errors
Lab products found in correlation
Octreotide, by Novartis (1 mentions)
Flutamide, by Merck Group (1 mentions)
Tin protoporphyrin snpp, by Merck Group (1 mentions)
Ne per nuclear and cytoplasmic extraction kit, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Cd86 pe, by Thermo Fisher Scientific (1 mentions)
Recombinant gm csf and il 4, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharide lps derived from escherichia coli 026 b6, by Merck Group (1 mentions)
Tin protoporphyrin 9 snpp, by Merck Group (1 mentions)
Mhcii fitc, by Thermo Fisher Scientific (1 mentions)
Cd80 fitc, by Thermo Fisher Scientific (1 mentions)
Goat anti rabbit igg horseradish peroxidase hrp, by Bioworld Technology (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
5 protocols using cobalt protoporphyrin copp
1
Reagents for HO-1 Antibody Experiments
2
Flutamide Treatment of HepG2 Cells
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HepG2 cells were obtained from Shanghai Cell Line Bank (Shanghai, China) and routinely cultured in MEM supplemented with 10% FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin. Cell cultures were grown at 37°C in humidified incubators containing an atmosphere of 5% CO2. All cells used in our experiments were up to passage 20. Cells were treated with flutamide (Sigma-Aldrich, USA) for 24 h at various concentrations as indicated. In some experiments, cells were pretreated with or without 20 μM tin protoporphyrin (Snpp, Sigma-Aldrich) or 10 μM cobalt protoporphyrin (Copp, Sigma-Aldrich) for 1 h followed by treatment with 50 μM flutamide.
3
Angiotensinogen Overexpression Impacts IRPTC Cells
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The IRPTC cell line11 (link),18 (link) and an IRPTC stable clone that has been stably transfected with the control plasmid pRC/RSV (designated as ‘pRSV-IRPTC’) or with a plasmid pRC/RSV containing the rat angiotensinogen cDNA (designated as ‘pRSV/rAgt-IRPTC’)13 (link),19 (link) were employed for our in vitro studies. We have previously reported that as compared to naive IRPTC and pRSV-IRPTC, pRSV/rAgt-IRPTC express significantly high amounts of rat angiotensinogen mRNA and protein as well as significantly higher amounts of Ang II secreted into the culture medium.13 (link),19 (link) All in vitro studies were performed in the normal 150 nM NaCl final concentration with an osmolarity of 415 mOsm/kg in normal glucose (5 mM D-glucose) DMEM as reported by Bouley et al.12 (link)Nuclear protein and cytosolic protein extracts were prepared using the NE-PER nuclear and cytoplasmic extraction kit (Thermo Scientific, Burlington, ON, Canada).18 (link) Cobalt protoporphyrin (CoPP, an activator of HO-1 expression) was purchased from Sigma-Aldrich Canada (Oakville, ON, Canada).
4
Characterization of Dendritic Cell Activation
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Fetal bovine serum (FBS) was purchased from Hyclone (Thermo, Melbourne, Australia). RPMI1640 medium (no phenol red), penicillin and streptomycin were purchased from Invitrogen (Grand Island, NY, USA). Recombinant GM-CSF and IL-4 were from Peprotech (Rocky Hill, NJ). Lipopolysaccharide (LPS) derived from Escherichia coli 026:B6 was from Sigma (St. Louis, MO). FITC-CD80, PE-CD86, FITC-MHCII or respective isotypes were from eBioscience (San Diego, CA, USA). Rabbit anti-mouse heme oxygenase 1(HO-1), rabbit anti-mouse β-actin and goat anti-rabbit IgG-horseradish peroxidase (HRP) were from Bioworld (St. Louis Park, MN, USA). Micro BCA protein assay kit was from Pierce (Rockford, IL). Cobalt protoporphyrin (CoPP), an inducer of HO-1 [38 (link)], and tin protoporphyrin IX (SnPP), an inhibitor of HO-1 activity [38 (link)], were from Sigma. Synthetic BP5 was purchased from Biotech Bioscience and Technology Co., Ltd (Shanghai, China). The purity of the synthetic peptide was > 98% by RP-HPLC. The sequence of the synthetic peptide was examined by electrospray ionisation tandem mass spectrometry (ESI-MS/MS). It was also tested the LPS contamination using the E-Toxate Limulus LPS detection kit (Sigma), which is sensitive to 0.05 to 0.1 endotoxin units/ml. Only uncontaminated preparations were used.
5
Cobalt Protoporphyrin Dissolution Protocol
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Cobalt protoporphyrin (CoPP) was obtained from Sigma Chemical Company (St. Louis, MO, USA) and dissolved in distilled water in dark tubes. Hydrochloric acid (HCl) was purchased from Sigma Aldrich fine chemicals (Cat #, H1758).
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