For decontamination of the PCR master mix, 8-methoxypsoralen was dissolved in dimethylsulfoxide (DMSO) (Sigma-Aldrich, St. Louis, MO, USA). For decontamination of the real-time PCR master mix reagents, 25 µg/mL of 8-methoxypsoralen (Sigma-Aldrich) and 10 min of exposure to UV irradiation at 366 nm over a distance of three cm were applied30 (link). Components included in the Sepsis@Quick kit were also decontaminated by UV irradiation at 280 nm over a distance shorter than five cm30 (link). Working places, as well as all accessories and tools, routinely undergo weekly decontamination by spraying with DNA-ExitusPlus™ solution (AppliChem, Darmstadt, Germany).
8 methoxypsoralen
Manufactured by Merck Group
Sourced in United States, Germany
8-methoxypsoralen is a chemical compound used in various laboratory applications. It is a naturally occurring substance derived from plants. The core function of 8-methoxypsoralen is to act as a photosensitizing agent, which means it can absorb ultraviolet light and transfer the energy to other molecules, inducing chemical reactions. This property makes it useful in certain experimental and analytical procedures.
Automatically generated - may contain errors
Lab products found in correlation
Omeprazole, by Merck Group (3 mentions)
Coumarin, by Merck Group (3 mentions)
Dna exitusplus solution, by AppliChem (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Clomethiazole, by Merck Group (2 mentions)
Quinidine, by Merck Group (2 mentions)
Sulfaphenazole, by Merck Group (2 mentions)
Furafylline, by Merck Group (2 mentions)
Ketoconazole, by Merck Group (2 mentions)
D glucose 6 phosphate, by Merck Group (2 mentions)
Glucose 6 phosphate dehydrogenase, by Merck Group (2 mentions)
Isopimpinellin, by Merck Group (2 mentions)
Oxypeucedanin, by Merck Group (2 mentions)
Oxypeucedanin hydrate, by Merck Group (2 mentions)
Nobiletin, by Merck Group (2 mentions)
Isomeranzin, by Merck Group (2 mentions)
Sinensetin, by Merck Group (2 mentions)
Tangeretin, by Merck Group (2 mentions)
5 geranyloxy 7 methoxycoumarin, by Merck Group (2 mentions)
Tetra o methylscutellarein, by Merck Group (2 mentions)
15 protocols using 8 methoxypsoralen
1
Decontamination of PCR Reagents
2
Decontamination of PCR Reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primers and labeled probes (Integrated DNA Technologies, Singapore) used are listed in Additional file 1 : Tables S2 and S3. All primers and probes were available in stocks of 100 pmol/μl in 25 mM Tris–HCl and treated with 1 U DNase/20 μl reaction volume for 15 min at 37 °C. Subsequently, the DNAse was inactivated by heating at 95 °C for 40 min. After inactivation, the stock was aliquoted into volumes of 10 μl and stored at −20 °C [16 (link)]. For decontamination of the PCR mastermix, 8-methoxypsoralen was dissolved in dimethylsulfoxide (DMSO) (Sigma, Germany). For decontamination of PCR water and the real-time PCR mastermix reagents, 25 μg/mL of 8-methoxypsoralen (Sigma, Germany) and 10 min exposure to 366 nm UV irradiation over a distance of 3 cm were applied. The MCLB-1 buffer was also decontaminated by UV irradiation at 280 nm over a distance shorter than 5 cm [16 (link)]. Working places and all accessories and tools in our laboratory are decontaminated weekly by spraying with DNA-ExitusPlus™ solution (AppliChem, Germany).
3
Decontamination Procedures for PCR Reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primers and labelled probes (Integrated DNA Technologies, Singapore) used are listed in Table 1. All primers and probes were treated with 1 U DNase in a 20 ml reaction volume for 15 min at 37 C. Subsequently, DNAse was inactivated by heating at 95 C for 40 min (Klaschik et al., 2002) . For decontamination of the PCR master mix, 8-methoxypsoralen was dissolved in dimethylsulfoxide (DMSO) (Sigma-Aldrich, St. Louis, MO, USA, Germany). For decontamination of real-time PCR master mix reagents, 25 mg/mL of 8-methoxypsoralen (Sigma-Aldrich) and 10 min of exposure to 366 nm UV irradiation over a distance of three cm were applied. The MCLB-1 buffer was decontaminated by UV irradiation at 280 nm over a distance shorter than five cm (Klaschik et al., 2002) . Working places as well as all accessories and tools routinely undergo weekly decontamination by spraying with DNA-Exitus-Plus TM solution (AppliChem, Darmstadt, Germany).
4
Xanthotoxol Cytochrome P450 Inhibition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Xanthotoxol (purity > 98%) was purchased from Sichuan Weikeqi Biotechnology Co. Ltd. (Sichuan, China). Paclitaxel, 1-aminobenzotriazole (ABT), phenacetin, sulfaphenazole, chlorzoxazone, quinidine, clomethiazole, furafylline, 8-methoxypsoralen, coumarin, diclofenac, quercetin, dextromethorphan, ketoconazole, testosterone, S-mephenytoin, omeprazole, glucose-6-phosphate dehydrogenase, NADP+, and D-glucose-6-phosphate were obtained from Sigma-Aldrich (St. Louis, MO, USA). All other reagents were the highest purity commercially available or HPLC grade.
5
Human Liver Microsome Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BFT and BF were purchased from Shanghai Winherb Medical Technology Company (Shanghai, China). ABT, furafylline, sulfaphenazole, clomethiazole, omeprazole, 8-methoxypsoralen, ticlopidine, CYP3cide, glucose-6-phosphate dehydrogenase, D -glucose-6-phosphate, and NADP+ were obtained from Sigma (St. Louis, MO, United States). Montelukast, quinidine, ketoconazole was purchased from Jianglai Biotechnology Co., Ltd. (Shanghai, China). The pooled HLMs (from 50 donors, lot no. X008067) were obtained from BioreclamationIVT (Baltimore, MD, United States). A panel of baculovirus expressed human P450s (CYP1A1, 1A2, 2A6, 2A13, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 3A4, 3A5, 4F2, and 4F3), co-expressing NADPH-CYP reductase and cytochrome b5 were obtained from BD Gentest Corp (Woburn, MA, United States). All chemicals and solvents were of analytical grade.
6
Selective Fluorescence Probes for CYP2A6 and GGT
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To demonstrate substrate selectivity of coumarin for CYP2A6, and gGlu-HMRG for GGT, 10mM of 8-Methoxypsoralen (M3501, Sigma-Aldrich), a known inhibitor of CYP2A6 (Pelkonen et al., 2000 (link)), or 50μM of GGsTop (4452, Tocris Bioscience, Bristol, UK), a known inhibitor of GGT (Han et al., 2007 (link)) were applied to the mouse nasal mucosa 5 min before administrating activatable fluorescence probes, respectively.
7
Recombinant CYP2A6 Variant Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Luria-Bertani and Teriffic broth media were purchased from Invitrogen Corporation (Carlsbad, CA, US). Tris base was acquired from Promega (Madison, WI, USA) while acetonitrile and hydrochloride acid were from Fisher Scientific (Pittsburgh, PA, USA). Coumarin, 7-hydroxyCoumarin, 8-methoxypsoralen, nicotinamide adenine phosphate (NADP+), glucose-6-phosphate dehydrogenase (G6PDH), glucose-6-phosphate (G6P), dimethyl sulfoxide (DMSO) and magnesium chloride were obtained from Sigma-Aldrich (St. Louis, MI, US). The collection of Escherichia coli (E. coli) bacterial stocks harbouring the plasmids of the wild type CYP2A6 (pCW-CYP2A6*1), all four individual variants of CYP2A6 (pCW-CYP2A6*15, pCW-CYP2A6*16, pCW-CYP2A6*21 and pCW-CYP2A6*22) and pACYC-oxidoreductase (pACYC-OxR) was previously constructed and prepared in our laboratory [9] (link).
8
Comprehensive Protocol for Photochemical Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
2,7-Dichlorodi-hydrofluorescein diacetate (DCFH-DA) was purchased from Bu Bai Chemical Technology Co., Ltd. (Shanghai, China). 4-tert-Buty-4′-methoxydibenzoylmethane (also known as avobenzene, Parsol 1789, etc.) was purchased from King Jack Int., Hong Kong. The cream substrate was a kind gift from Department of Dermatology, Huashan Hospital (Shanghai, China). Neutral Red (NR) was purchased from Shanghai Source Leaves Biological Co., Ltd. (Shanghai, China). 8-Methoxypsoralen was purchased from Sigma (St Louis, MO, USA). All other regents used were of analytical grade. The synthetic routes of CAB and ACB have been described by our group (Scheme 1 ).11,22 (link)
9
Live Imaging of Damaged Chromatin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HAP1 cells expressing the indicated GFP-tagged proteins were grown on Nunc Lab Tek II chambered cover glasses (Thermo Fisher Scientific). Protein expression was induced using doxycycline (2 μg/ml) for 24 hours. Next, cells were pretreated with 10 μM 8-methoxypsoralen (Sigma-Aldrich) for 1 hour before UVA microirradiation using a 355-nm laser (CNI, AO-S-355-40 mW, power: 10 μW, power density: 50 to 100 W/cm2) focused through a 63× oil objective with numerical aperture of 1.4 to yield a stripe size of 20 × 0.5 μm. The imaging was performed on a Leica STELLARIS 8 confocal microscope, and a white light laser at 488 nm was used (10% power). Stripes were imaged for 10 hours after UVA microirradiation.
10
Coumarin Oxidation Kinetics in Liver
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In vitro oxidation rates of coumarin catalyzed by liver preparations and hepatocytes from rats or humans were measured using liquid chromatography (LC) with fluorescence and ultraviolet detectors (Yamazaki et al., 1999) with an analytical reversed-phase column. Briefly, coumarin (10 µM, unless otherwise specified) was incubated with liver microsomes (0.50 mg protein/mL) in the presence or absence of pooled liver cytosolic fractions (1.0 mg/mL) or liver S9 fractions (1.0 mg/mL) and an NADPH-generating system with or without uri- dine diphosphate glucuronic acid (UDP-GA, 2.0 mM) or 3'-phosphoadenosine-5'-phosphosulfate (0.20 mM) in a total of 0.50 mL of 100 mM potassium phosphate buffer (pH 7.4) at 37°C for 15 min. The effects of furafylline/ α-naphthoflavone, 8-methoxypsoralen, 4-methylpyrazole, and ketoconazole (Sigma-Aldrich, Milwaukee, WI, USA), P450 1A2, 2A6, 2E1, 3A inhibitors (Yamazaki et al., 1992; Okubo et al., 2016) , respectively, on coumarin oxidation activities were also investigated in human liver microsomes. Coumarin (0.20, 2.0, and 20 µM) was incubated in the culture medium for hepatocytes (5 × 10 5 cells/well) at 37°C for 2 hr at 95% humidity and 5% CO 2 in a 0.50-mL reaction mixture. After test solutions were treated with an equal volume of acetonitrile, the aqueous supernatant was centrifuged at 2 × 10 3 g for 10 min at 4°C and analyzed.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!