Ovarian carcinoma SKOV3 cell lines were obtained from the Shanghai Institute of Cell Library (Shanghai, China). The small interfering RNA (siRNA) was designed and constructed as a vector (plasmid without vector and bacterial strain containing this plasmid were all produced and identified by Nanjing Moji Biological Technology Development Co. Ltd., Nanjing, China). The main reagents used were: CCK-8, IL-8, RPMI-1640 basal culture medium, double-antibody and fetal bovine serum (FBS) (both from Gibco, Grand Island, NY, USA), acrylamide, methylene acrylamide, Tris-base, ammonium peroxydisulfate, TEMED, glycine and SDS (all from Biosharp, Hefei, China). The PVDF membrane, and chemiluminescent liquid were purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA).
Tris base
Manufactured by Biosharp
Sourced in China
Tris-base is a chemical compound used as a buffer in various laboratory applications. It is a white crystalline solid that is soluble in water and is commonly used to maintain a stable pH in biological and biochemical experiments.
Automatically generated - may contain errors
Lab products found in correlation
Glycine, by Biosharp (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Sodium dodecyl sulfate (sds), by Biosharp (2 mentions)
Double antibody, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Thermo Fisher Scientific (1 mentions)
Cck 8, by Thermo Fisher Scientific (1 mentions)
Acrylamide, by Biosharp (1 mentions)
Bromophenol blue, by Thermo Fisher Scientific (1 mentions)
Urea, by Bio-Rad (1 mentions)
Agarose, by Biosharp (1 mentions)
Protease inhibitory cocktail, by Merck Group (1 mentions)
Glycerol, by Biosharp (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Collagenase type 1, by Merck Group (1 mentions)
Fast sybr green master mix, by Toyobo (1 mentions)
Tris hcl, by Bio-Rad (1 mentions)
Trizol reagent, by Takara Bio (1 mentions)
Thiourea, by Merck Group (1 mentions)
Irbesartan, by Merck Group (1 mentions)
Ugt1a9, by BD (1 mentions)
5 protocols using tris base
1
SKOV3 Cell Line Knockdown Protocol
2
Two-dimensional Proteomic Analysis of Cell Lines
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Dulbecco’s Modified Eagle’s Medium (DMEM) was purchased from Gibco (Waltham, MA, USA), and fetal bovine serum (FBS) was also purchased from Gibco. Urea, CHAPS zwitterionic detergent (CHAPS), dithiothreitol (DTT), immobilized pH gradient (IPG) buffer, Tris-HCl, and IPG strips (17 cm, pH 4–7, linear) were purchased from Bio-Rad (Hercules, CA, USA). Collagenase (Type I), protease inhibitory cocktail, and thioUrea were purchased from Sigma-Aldrich (St Louis, MO, USA). Sodium dodecyl sulfate (SDS), glycerol, Tris base, agarose, glycine, and Coomassie R-250 were purchased from Biosharp (Hefei, China). Iodoacetamide was purchased from TCI (Portland, OR, USA). Bromophenol blue was purchased from Fisher Scientific (Pittsburgh, PA, USA). TRIzol reagent and reverse transcription kit were purchased from Takara (Shiga, Japan). Fast SYBR Green Master Mix was purchased from Toyobo (Osaka, Japan).
3
Microsome-mediated Glucuronidation Assay
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Ib, Ib glucuronide and irbesartan (internal standard, IS) were supplied by the Department of Medicinal Chemistry of China Pharmaceutical University, Nanjing, China. UPDGA (63700-19-6), d -saccharic acid 1,4-lactone (61278-30-6), alamethicin (27061-78-5), BSA (9048-46-8), NADP (53-59-8), G-6-P (3671-99-6), and PDH (9001-40-5) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Tris-Base were purchased from Biosharp (Seoul, South Korea).
Mixed Gender Pooled Human Liver Microsomes (HLM, from 25 subjects of 25–89 years age), Mixed Gender Pooled Cynomolgus monkey Liver Microsomes (MKLM, from 14 subjects of 4–5 years age), Mixed Gender Pooled Beagle Dog Liver Microsomes (DLM, from 7 subjects of 11 weeks age), Mixed Gender Pooled Sprague-Dawley Rat Liver Microsomes (RLM, from 100 subjects of 2–3 months age) and Mixed Gender Pooled CD1 Mice Liver Microsomes (MLM, from 500 subjects of 5–8 weeks age) were purchased from the Research Institute for Liver Diseases (Shanghai) Co. Ltd. (Shanghai, China). Supersome preparations containing recombinant UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A7, UGT1A8, UGT1A9, UGT1A10, UGT2B4, UGT2B7, UGT2B15 or UGT2B17 (expressed in baculovirus-infected insect cells) and UGT Inset Cell Control SupersomesTM were purchased from BD Biosciences (San Jose, CA, USA). All other chemicals and solvents used were of the highest quality and analytical grade or higher.
Mixed Gender Pooled Human Liver Microsomes (HLM, from 25 subjects of 25–89 years age), Mixed Gender Pooled Cynomolgus monkey Liver Microsomes (MKLM, from 14 subjects of 4–5 years age), Mixed Gender Pooled Beagle Dog Liver Microsomes (DLM, from 7 subjects of 11 weeks age), Mixed Gender Pooled Sprague-Dawley Rat Liver Microsomes (RLM, from 100 subjects of 2–3 months age) and Mixed Gender Pooled CD1 Mice Liver Microsomes (MLM, from 500 subjects of 5–8 weeks age) were purchased from the Research Institute for Liver Diseases (Shanghai) Co. Ltd. (Shanghai, China). Supersome preparations containing recombinant UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A7, UGT1A8, UGT1A9, UGT1A10, UGT2B4, UGT2B7, UGT2B15 or UGT2B17 (expressed in baculovirus-infected insect cells) and UGT Inset Cell Control SupersomesTM were purchased from BD Biosciences (San Jose, CA, USA). All other chemicals and solvents used were of the highest quality and analytical grade or higher.
4
Synergistic Anti-cancer Effects of GSK126 and Gefitinib
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SRB assays were performed to detect cell proliferation. Cancer cells in logarithmic growth phase were seeded into 96-well plates at 1×104 cells/well and cultured overnight before treatment with GSK126 and/or Gefitinib at the indicated concentrations. After 48 hours of exposure, the medium was removed and cells were incubated with 50 µL of cold 10% trichloroacetic acid (Sigma-Aldrich) for 30 minutes at 4°C. The cells were then washed with phosphate-buffered saline (PBS) and stained with 0.4% SRB for 30 minutes at room temperature. After rinsed with 1% acetic acid and dried, cells were incubated with 100 µL Tris-base (Biosharp, Hefei, China) at 10 mM and violently shaken for 10 minutes. Absorbance was measured at 510 nm using a Spectramax M5 spectrophotometer (Molecular Devices, San Jose, CA, USA). The combination index (CI) of GSK126 and Gefinitib was calculated and their synergy quantification was performed using the Chou-Talalay method as described previously.20 (link) CI <1, CI =1, and CI >1 indicate synergism, additive effect, and antagonism in drug combinations, respectively.
5
Reagents for Oxidative Stress Assay
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The H
2O
2 solution (30%) was purchased from Jiangsu Yong Hua Chemical Technology Co., Ltd. (Changshu, Jiangsu, China), GEM from Saen Chemical Technology Co., Ltd. (Shanghai, China), trichloroacetic acid from Aladdin (Cat. #T104257, Shanghai, China), Tris-base from Biosharp (Cat. #BS083, Hefei, Anhui, China), and chromatographic methanol from Anhui Shi Lian Special Solvent Co., Ltd. (Anqing, Anhui, China). Ultrapure water was prepared by the ultrapure water system, other reagents were of analytical purity.
2O
2 solution (30%) was purchased from Jiangsu Yong Hua Chemical Technology Co., Ltd. (Changshu, Jiangsu, China), GEM from Saen Chemical Technology Co., Ltd. (Shanghai, China), trichloroacetic acid from Aladdin (Cat. #T104257, Shanghai, China), Tris-base from Biosharp (Cat. #BS083, Hefei, Anhui, China), and chromatographic methanol from Anhui Shi Lian Special Solvent Co., Ltd. (Anqing, Anhui, China). Ultrapure water was prepared by the ultrapure water system, other reagents were of analytical purity.
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