Biliverdin hydrochloride (Frontier Scientific, Logan, UT, USA) was dissolved in 0.2 mol/L NaOH, and the solution was adjusted to a final pH of 7.4 with hydrogen chloride. L-glutathione reduced (GSH) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Male Sprague-Dawley rats, weighing 200–230 g, were purchased from Jiangnan University Animal Center and housed in a room with a 12/12-h light/dark cycle. Rats had ad libitum access to food and water. Experiments were carried out according to protocols approved by the Animal Care and Use Committee of Jiangnan University.
Biliverdin hydrochloride
Manufactured by Frontier Scientific
Sourced in United States
Biliverdin hydrochloride is a chemical compound with the formula C33H36N4O6·HCl. It is a green crystalline solid that is used as a reagent in biochemical and analytical applications.
Automatically generated - may contain errors
Lab products found in correlation
Tricarbonyldichlororuthenium 2 corm2, by Merck Group (2 mentions)
Feso4, by Merck Group (2 mentions)
Tin protoporphyrin 9 snpp, by Frontier Scientific (2 mentions)
L glutathione reduced gsh, by Merck Group (1 mentions)
Hiload 26 600 superdex200 pg, by GE Healthcare (1 mentions)
Histrap column, by GE Healthcare (1 mentions)
8 protocols using biliverdin hydrochloride
1
Biliverdin Hydrochloride and Glutathione Rat Study
2
In Vivo and In Vitro Tin Protoporphyrin IX Protocol
3
Purification of Phytochromes and Response Regulators
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All DrBphP variants and the response regulators were expressed in Escherichia coli strain BL21 (DE3) overnight at 20–24 °C. After cell lysis with EmulsiFlex®, a molar excess of biliverdin hydrochloride (Frontier Scientific) was added to the phytochrome samples and incubated overnight on ice. No external biliverdin was added to the cell lysate in response regulator purifications. The His6-tagged proteins were purified with NiNTA affinity purification using HisTrap™ columns (GE Healthcare), followed by size-exclusion chromatography (HiLoad™ 26/600 Superdex™ 200 pg, GE Healthcare) in buffer (30 mM Tris, pH 8.0)76 (link). Agp1 and its D529H mutant were expressed in NEB Express® IqE. coli strain (New England Biolabs). The purification protocol was identical to other samples with a couple of exceptions: Protease inhibitor mix (ROCHE) and 0.5 mM TCEP were included in the sample before lysis, and affinity purification was conducted in (30 mM Tris/HCl, 150 mM NaCl, 1 mM TCEP) and varying imidazole concentration (5–500 mM). All purified protein samples were concentrated to 25–30 mg/ml in (30 mM Tris/HCl, pH 8.0) and flash-frozen.
4
Biliverdin Supplementation for Transgenic C. elegans
5
In Vivo and In Vitro Tin Protoporphyrin IX Protocol
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For all in vivo tin protoporphyrin IX (SnPP) (Frontier Scientific, Logan, UT) treatments, mice were intraperitoneal injected daily (5 mg/kg/mouse in 200ul of PBS) starting 28 days post-infection. In vitro, SnPP was added to culture medium for at a concentration of 1μM. In in vitro experiments, biliverdin hydrochloride (Frontier Scientific, Logan, UT), Tricarbonyldichlororuthenium II (CORM2) or FeSO4 (both from Sigma-Aldrich, Saint Louis, MO), were added to cell culture medium all at 5μM.
6
Worm Culture with Biliverdin and Fluorescent Reporters
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Worms were grown at 20 ºC in constant darkness on agar plates containing nematode growth medium and fed the OP50 derivative bacterial strain DA837 (45 (link)), which was supplemented with 100 μm biliverdin hydrochloride (Frontier Scientific). The following strains were used: N2 (Bristol), SJU16 = stjEx9[Punc-17:IlaC22:SL2:dsRed; Pmyo-2:gfp], SJU17 = stjEx10[Punc-17:IlaC22:SL2:dsRed; Pmyo-2:gfp], SJU159 = stjEx112[Punc-17:PaaC:SL2:dsRed; Pmyo-2:mCherry] and SJU161 = stjEx112[Punc-17:PaaC:SL2:dsRed; Pmyo-2:mCherry].
7
Purification and Recrystallization of UCB
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All chemicals were from Sigma (MO, USA), except for UCB and biliverdin hydrochloride (both from Frontier Scientific, UT, USA). Before use, UCB was purified and re-crystalized as described previously13 (link).
8
Biliverdin HPLC Quantification Protocol
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Biliverdin hydrochloride was obtained from Frontier Scientific (Utah, USA). All other reagents were obtained from Sigma-Aldrich, unless otherwise stated. BV standards were stored at −30 °C (in DMSO) and biological samples were stored at −80 °C. Prior to injection, 40 µL of either standard or sample was added to 120 µL of mobile phase (50:50 20 mM ammonium acetate in 100% HPLC grade methanol: 20 mM ammonium acetate in HPLC grade H2O), except for duodenal samples where 120 µL of pure methanol (duodenum) was used instead of mobile phase in the same ratios.
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