MN (purity > 98%) and HK (purity > 98%) were purchased from ChemFaces (Wuhan, PRC). Ko143 (purity 96%) was obtained from Enzo Life Sciences, Inc. (Farmingdale, NY, USA). MXR (purity 99%) was obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Dimethyl sulfoxide (DMSO), formic acid, sodium dodecyl sulfate (SDS), 3-(4′,5′-dimethylthiazol-2′-yl)-2,5-diphenyltetrazolium bromide (MTT), butylparaben and triton X-100 were supplied by Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Fetal bovine serum (FBS) was obtained from Biological Industries Inc. (Kibbutz, Beit Haemek, Israel). Penicillin-Streptomycin-Glutamine, Dulbecco’s modified Eagle medium (DMEM), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, Hank’s buffered salt solution (HBSS)and trypsin/EDTA were purchased from Invitrogen (Grand Island, NY, USA). Antibodies against ABCG2 and GAPDH were purchased from GeneTex (San Antonio, TX, USA). Polyvinylidene fluoride transfer membranes (Immobilon P) and chemiluminescence (ECL) were obtained from Millipore Corp. (Bedford, MA, USA). 4′,6-diamidino-2-phenylindole (DAPI) staining solution and goat anti-rabbit IgG H&L (DyLight™ 488) antibody were purchased from Abcam (Cambridge, UK). Acetonitrile (ACN) and methanol with LC grade were obtained from Mallinckrodt Baker (Phillipsburg, NJ, USA). Milli-Q plus water (Bedford, MA, USA) was used throughout this study.
Hank s buffered salt solution hbss
Manufactured by Thermo Fisher Scientific
Sourced in United States, France
Hank's Buffered Salt Solution (HBSS) is a balanced salt solution commonly used in cell culture and biological research applications. It provides a physiologically relevant environment for the maintenance and handling of cells in vitro. The solution contains a carefully balanced combination of inorganic salts, glucose, and other components to support cell viability and function.
Automatically generated - may contain errors
Lab products found in correlation
Ko143, by Enzo Life Sciences (2 mentions)
Penicillin streptomycin glutamine (psg), by Thermo Fisher Scientific (2 mentions)
Dispase, by Thermo Fisher Scientific (2 mentions)
Collagenase, by Merck Group (2 mentions)
Dnase, by Merck Group (2 mentions)
Hepes, by Merck Group (2 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by GE Healthcare (2 mentions)
Phenylmethylsulfonyl fluoride pmsf, by Merck Group (2 mentions)
Cholesterol, by Thermo Fisher Scientific (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Celltox green cytotoxicity assay kit, by Promega (2 mentions)
Hepes buffer, by MP Biomedicals (2 mentions)
Diacetyl phosphate, by Thermo Fisher Scientific (2 mentions)
Animal endothelial cell medium, by Cell Biologics (2 mentions)
Murine endothelial growth factor, by Cell Biologics (2 mentions)
Fetal bovine serum (fbs), by Cell Biologics (2 mentions)
L glutamine, by Cell Biologics (2 mentions)
Antibiotic antimycotic, by Cell Biologics (2 mentions)
D luciferin fluorescence dye, by BD (2 mentions)
15 protocols using hank s buffered salt solution hbss
1
Evaluation of Anticancer Compound Purity
2
Warfarin-Resveratrol Interaction Assay
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(±)Warfarin sodium (purity 98%), caffeic acid (purity 98%), formic acid, 3-(40,50-dimethylthiazol-20-yl)-2,5-diphenyltetrazolium bromide (MTT) and sodium dodecyl sulfate were purchased from Sigma-Aldrich Chemical Co. (MO, U.S.A.). R-warfarin (purity 98%) and S-warfarin (purity 98%) were obtained from Toronto Research Chemicals, Inc. (Ontario, Canada). Ko143 (purity 96%) was obtained from Enzo Life Sciences, Inc. (NY, U.S.A) Fetal bovine serum, Dulbecco’s modified Eagle medium, penicillin–streptomycin-glutamine and Hank’s buffered salt solution (HBSS) were obtained from Invitrogen (CA, U.S.A). Resveratrol (purity > 99%) was obtained from TCI (Tokyo, Japan). Trans-resveratrol capsules were purchased from Doctor’s BEST (CA, U.S.A.). Acetonitrile and methanol of LC–MS grade were obtained from J.T. Baker Inc. (PA, U.S.A.).
3
Isolation of Human Prostate Epithelial Cells
4
Isolation of Human Prostate Epithelial Cells
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Human prostate digestion and isolation of epithelial cells were performed as described previously [24 (link)]. Briefly, fresh human prostate tissue, harvested from radical prostatectomy surgery stored until digestion in static preservation solution (SPS-1™) (Organ Recovery Systems, Buffalo, NY) at 4°C was obtained from the Pathology Resource Network at RPCI. Enzymatic tissue digestion with a mixture of: dispase (2.4 U/ml) (Invitrogen, Carlsbad, CA); DNase (0.01% w/v) (Sigma Aldrich, St. Louis, MO); and collagenase (2.8% w/v) (Sigma Aldrich, St. Louis, MO) followed by single cell suspension in Hanks buffered salt solution (HBSS) (Invitrogen, Carlsbad, CA) supplemented with 5% FBS was prepared as described previously [24 (link)].
5
Isolation and Characterization of Breast Tissue-Derived Mesenchymal Stem Cells
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Human Healthy Breast Tissue-Mesenchymal Stem Cells (hHBT-MSCs) were obtained from breast reduction surgeries, whereas Human Malignant Breast Tissue-Mesenchymal Stem Cells (hMBT-MSCs) were obtained from tumor removal surgeries. In a sterile environment (class II "Laminar Cabinet), breast tissues from healthy and cancer patients were placed in a petri dish and washed with 5% antibiotic Hank's Buffered Salt Solution (HBSS) (Gibco Invitrogen, Grand Island, USA) and cut into small pieces with scissors and treated with the enzyme collagenase type II (Gibco Invitrogen, Grand Island, USA). After washing, the supernatant was removed and the cells were cultured with DMEM/F12 (Hyclone, USA) medium containing Fetal Bovine Serum (FBS), antibiotics, insulin and hydrocortisone at 37°C in a humid atmosphere with 5% CO2. On the third day, non-adherent cells were removed and fresh medium was added. When the bottom of the ask was approximately 70% covered with cells, passage was performed. At the third passage (P3), characterization of the stromal cells was performed.
The breast cancer cell line MDA-MB-231 (ATCC) was also grown and propagated in medium enriched with serum, antibiotics, insulin and hydrocortisone (DMEM / F12), which showed good proliferation.
The breast cancer cell line MDA-MB-231 (ATCC) was also grown and propagated in medium enriched with serum, antibiotics, insulin and hydrocortisone (DMEM / F12), which showed good proliferation.
6
Characterization of Transporter Substrates
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Ipratropium bromide, IBC, IBC-d6, l -noradrenaline, ritodrine hydrochloride, sumatriptan-d6, thiamine-d3 hydrochloride, and trospium chloride were obtained from Santa Cruz Biotechnology. Ranitidine-d6 and trospium-d8 were obtained from Toronto Research Chemicals, ASP+ was obtained from Life Technologies, and buformin hydrochloride was obtained from Wako Chemicals. Radiolabeled MPP+ (N-[methyl-3H], 80 Ci/mmol) and TEA+ ([ethyl-1-14C]; 55 mCi/mol) were obtained from Hartmann Analytic. All other compounds tested as substrates were obtained from Sigma–Aldrich. All chemicals used in this study were purchased from commercial sources and had purities of 97% or higher.
Dulbecco's modified Eagle's medium, Hank's buffered salt solution (HBSS), and additives used for cell culturing were obtained from Life Technologies. Poly-d -lysine (1–5 kDa), Hepes, bicinchoninic acid, and copper sulfate pentahydrate were obtained from Sigma–Aldrich. Twelve-well plates were obtained from Starlab, and tissue culture flasks were from Sarstedt. Acetonitrile and methanol in LC–MS/MS grade were obtained from LGC Standards, and formic acid (LC–MS/MS grade) and sodium chloride were obtained from Merck. SDS (UltraPure) was obtained from AppliChem.
Dulbecco's modified Eagle's medium, Hank's buffered salt solution (HBSS), and additives used for cell culturing were obtained from Life Technologies. Poly-
7
Cannabinoid and Vanilloid Signaling Pathways
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Cannabidiol (CBD) (1 mg/mL in methanol), tranilast (TNL), capsaicin (CAP), capsazepine (CPZ), and GSK1016790A (GSK) were all analytical grade and purchased from Sigma-Aldrich (Saint-Quentin-Fallavier, France). CaCl2, MgSO4, NaCl, NaH2PO4, NaHCO3, KH2PO4, and KCl were purchased from Merck (Fontenay-sous-Bois, France).
RNA extraction kits were bought from Qiagen (Courtaboeuf, France). Primers, RT-PCR reagents and Lipofectamine® RNAi MAX transfection reagent were obtained from Invitrogen Life Technologies (Cergy-Pontoise, France). The Power SYBR Green PCR Master Mix was purchased from Applied Biosystems (Applied BiosystemsTM, France). All other reagents and chemicals were purchased from Sigma.
Dulbecco’s modified Eagle’s medium (DMEM), DMEM/F12 + Glutamax, Hank’s buffered salt solution (HBSS), HEPES and Penicillin/Streptomycine were from Life Technologies (Cergy-Pontoise, France). fetal bovine serum (FBS) was from GE Healthcare Life Science. Liberase DL, bovine serum albumin (BSA) (cat. # A7906), Dextran (cat. # 31390), DNase I (cat. # DN25), Puromycine and b-FGF were from Sigma-Aldrich. plasma derived bovine serum (PDBS) was from First Link (Wolverhampton, United Kingdom).
RNA extraction kits were bought from Qiagen (Courtaboeuf, France). Primers, RT-PCR reagents and Lipofectamine® RNAi MAX transfection reagent were obtained from Invitrogen Life Technologies (Cergy-Pontoise, France). The Power SYBR Green PCR Master Mix was purchased from Applied Biosystems (Applied BiosystemsTM, France). All other reagents and chemicals were purchased from Sigma.
Dulbecco’s modified Eagle’s medium (DMEM), DMEM/F12 + Glutamax, Hank’s buffered salt solution (HBSS), HEPES and Penicillin/Streptomycine were from Life Technologies (Cergy-Pontoise, France). fetal bovine serum (FBS) was from GE Healthcare Life Science. Liberase DL, bovine serum albumin (BSA) (cat. # A7906), Dextran (cat. # 31390), DNase I (cat. # DN25), Puromycine and b-FGF were from Sigma-Aldrich. plasma derived bovine serum (PDBS) was from First Link (Wolverhampton, United Kingdom).
8
Cytotoxicity Assay Protocol
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Sodium arsenite (Merck, Germany), cadmium chloride (Sigma-Aldrich, USA) and mercury chloride (BDH, UK) were prepared in deionised water. BBL™ FTA hemagglutination buffer was obtained from BD (France) and used to prepare phosphate-buffered saline (PBS).
Hank's Buffered Salt Solution (HBSS) was purchased from Life Technologies (South Africa). Penicillin (10,000 IU) and streptomycin (10 mg/mL) solutions were obtained from BioWhittaker (Walkersville, USA). Acetyl-Asp-Glu-Val-Asp-7-amido-4-methylcoumarin (Ac-DEVD-AMC), ATP Assay Kit MAK135, 2,2'-azobis (2-methylpropionamide) dihydrochloride (AAPH), camptothecin, 3-[(3-chloamidopropyl)dimethylammonio]-1-propanesulfonate, 2',7'dihydrodichlorofluorescein diacetate (H2-DCF-DA), Dulbecco's Modified Eagle's Medium (DMEM), ethylenediaminetetraacetic acid (EDTA), HEPES, JC-1, monochlorobimane, nethylmaleimide, phenylmethylsulfonyl fluoride (PMSF), rotenone, saponin, staurosporine, sulforhodamine B (SRB), trypsin, Tris-base and trypan blue were obtained from Sigma-Aldrich (St. Louis, USA). β-Mercaptoethanol and trichloroacetic acid (TCA) was obtained from Merck Chemicals (Darmstadt, Germany). Foetal calf serum (FCS) was procured from PAA (Pasching, Austria).
Hank's Buffered Salt Solution (HBSS) was purchased from Life Technologies (South Africa). Penicillin (10,000 IU) and streptomycin (10 mg/mL) solutions were obtained from BioWhittaker (Walkersville, USA). Acetyl-Asp-Glu-Val-Asp-7-amido-4-methylcoumarin (Ac-DEVD-AMC), ATP Assay Kit MAK135, 2,2'-azobis (2-methylpropionamide) dihydrochloride (AAPH), camptothecin, 3-[(3-chloamidopropyl)dimethylammonio]-1-propanesulfonate, 2',7'dihydrodichlorofluorescein diacetate (H2-DCF-DA), Dulbecco's Modified Eagle's Medium (DMEM), ethylenediaminetetraacetic acid (EDTA), HEPES, JC-1, monochlorobimane, nethylmaleimide, phenylmethylsulfonyl fluoride (PMSF), rotenone, saponin, staurosporine, sulforhodamine B (SRB), trypsin, Tris-base and trypan blue were obtained from Sigma-Aldrich (St. Louis, USA). β-Mercaptoethanol and trichloroacetic acid (TCA) was obtained from Merck Chemicals (Darmstadt, Germany). Foetal calf serum (FCS) was procured from PAA (Pasching, Austria).
9
In Vitro BBB Endothelial Cell Assay
10
Evaluating SATA-8505 Activity Against USA300
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Approximately 106 CFUs of mid-exponential growth phase USA300 were added to 5 mL aliquots of BHI or TSB with SATA-8505 at varying MOI. Samples were incubated with shaking at 37°C, with bacterial concentration over time determined by plating on BHI (BD Biosciences) or blood (Thermo Fisher Scientific) agar. For the whole blood assay, 10mL blood from normal donors and/or CGD patients was obtained from the NIH Clinical Center Department of Transfusion Medicine and was added to blood culture bottles containing 30mL of TSB (Thermo Fisher Scientific) with SATA-8505 at varying MOI. In experiments using blood, control blood culture vials were injected with an equal volume of Hanks Buffered Salt Solution (HBSS) (Thermo Fisher Scientific). All human samples were obtained under permission of the Institutional Review Board (IRB) for the National Institute of Allergy and Infectious Disease (NIAID) and the National Institutes of Health (NIH) Clinical Center. All participants provided their written consent to the research protocol and IRB consent was obtained prior to blood collection.
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