n g nitro l arginine nla  (Millipore)


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    Millipore n g nitro l arginine nla
    N G Nitro L Arginine Nla, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Centrifugation:

    Article Title: The structure of avian polyomavirus reveals variably sized capsids, nonconserved inter-capsomere interactions, and a possible location of the minor capsid protein VP4
    Article Snippet: After removing cellular debris by low-speed centrifugation, the supernatant was loaded onto a two-step, CsCl density gradient (ρ = 1.3 and 1.4 g/ml). .. L-arginine has been used to reduce protein aggregation , and we treated purified APV with GP buffer plus 250 mM L-arginine (APV+R) by dialysis (D-tube Mini Dialyzers, MWCO 12-14 kDa, Novagen) to stabilize and de-aggregate particles.

    Filtration:

    Article Title: The structure of avian polyomavirus reveals variably sized capsids, nonconserved inter-capsomere interactions, and a possible location of the minor capsid protein VP4
    Article Snippet: L-arginine has been used to reduce protein aggregation , and we treated purified APV with GP buffer plus 250 mM L-arginine (APV+R) by dialysis (D-tube Mini Dialyzers, MWCO 12-14 kDa, Novagen) to stabilize and de-aggregate particles. .. The sample was subjected to centrifugal filtration (YM-100 centrifugal devices, Millipore, Billerica, Massachusetts, USA) for a final washing and concentrating step.

    Article Title: Biochemical enrichment and biophysical characterization of a taste receptor for L-arginine from the catfish, Ictalurus puntatus
    Article Snippet: The absolute enantiomer, L-arginine HCl, was purchased from Sigma, and the absolute enantiomer, D-arginine HCl, was a gift of the Ajinomoto Co., Tokyo, Japan. .. Sephacryl S-300 HR, High Range Gel Filtration Calibration Kits, and 1.0 ml Hitrap Q columns were purchased from Pharmacia Biotech (Piscataway, NJ).

    Cell Isolation:

    Article Title: Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function
    Article Snippet: Paragraph title: CD8 T cell isolation and in vitro activation ... Medium was supplemented with indicated concentrations of L-arginine (Sigma, #A5006) and/or L-ornithine (Sigma, #O2375) and cultured up to 72h before proceeding with intracellular cytokine staining.

    Modification:

    Article Title: Molecular Signature of CAID Syndrome: Noncanonical Roles of SGO1 in Regulation of TGF-β Signaling and Epigenomics
    Article Snippet: .. Cell culture and SILAC SILAC Dulbecco's modified Eagle medium without L-arginine and L-lysine (88364; Thermo Fisher Scientific, Waltham, MA) was supplemented with 13C6, 15N4 L-arginine (20104102; Silantes, München, Germany), 13C6, and 15N2 L-lysine (2111604102; Silantes), or unlabeled 12C614N4 L-arginine (A5006; Sigma Aldrich, St. Louis, MO) and 12C6L-lysine (L5501; Sigma Aldrich) to produce heavy and light SILAC media, respectively. .. L-proline (P5607; Sigma Aldrich) also was added to a final concentration of 100 μg/mL to prevent amino acid conversion.

    Article Title: Insert engineering and solubility screening improves recovery of virus-like particle subunits displaying hydrophobic epitopes
    Article Snippet: .. Modified DD IT1 capsomere recovered via SEC in buffer containing 50 m M l -arginine ( l -Arg) (≥98%, Sigma-Aldrich, ) at pH 8.5 was assembled by dialysis in standard assembly buffer containing 0.5 M (NH4 )2 SO4 , 20 m M Tris–base, 5% glycerol (v/v), 1 m M CaCl2 at pH 7.4, with or without l -Arg at 50 m M . .. Samples were placed on 200-mesh carbon-coated copper grids and left to settle for 90 s. Following two washes in water, grids were negative stained with 2% uranyl acetate for 60 s. TEM was carried out on a JEOL 1010 at 80 kV and images were cropped and adjusted for brightness using Adobe Photoshop (CS6; ).

    Incubation:

    Article Title: Essential Role for an M17 Leucine Aminopeptidase in Encystation of Acanthamoeba castellanii
    Article Snippet: Briefly, 10 μL of enzyme solution was added to 190 μL of assay buffer (50 mM Tris-HCl, 1 mM MnCl2 , pH 8.0) containing 10 μM Leu-AMC and incubated for 20 min at 37°C, and the release of fluorescence was measured at an excitation wavelength of 370 nm and an emission wavelength of 440 nm using a Gemini EM fluorescence microplate reader (Molecular Devices, Sunnyvale, CA, USA). .. The substrate specificity of AcLAPr was determined using Leu-AMC, l -arginine-AMC (Arg-AMC), and l -methionine-AMC (Met-AMC) (all from Sigma-Aldrich) as substrates.

    Article Title: Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function
    Article Snippet: The pellet was resuspended in 1 mL red blood cell lysis buffer (eBioscience, #00-4300-54) and incubated at room temperature for 1 min. .. Medium was supplemented with indicated concentrations of L-arginine (Sigma, #A5006) and/or L-ornithine (Sigma, #O2375) and cultured up to 72h before proceeding with intracellular cytokine staining.

    Isotopic Labeling:

    Article Title: Cell-specific Labeling Enzymes for Analysis of Cell–Cell Communication in Continuous Co-culture *
    Article Snippet: All cells were obtained from ATCC Manassas, VA (except KPC cells, a kind gift from Professor Owen Sansom, Glasgow) and were grown in DMEM (deficient for L -lysine and L -arginine) (DMP49, Caisson, Utah) supplemented with 10% (v/v) dialyzed FBS (Invitrogen), 0.3 m m L -arginine (A8094, Sigma) and/or L -lysine (L5501, Sigma), D -lysine (L5876, Sigma), and DAP (07036, Sigma). .. For SILAC isotopic labeling, cells were grown in 2.5 m m “medium” L -lysine (Isotec 616192, Sigma) (delta mass: 4.025107; delta average mass: 4.0246) or “heavy” L -lysine (Isotec 608041, Sigma) (delta mass: 8.014199; delta average mass: 7.9427).

    Activity Assay:

    Article Title: Essential Role for an M17 Leucine Aminopeptidase in Encystation of Acanthamoeba castellanii
    Article Snippet: The effects of an aminopeptidase inhibitor (bestatin; Sigma-Aldrich) and metal chelators (EDTA and 1,10-phenanthroline; Sigma-Aldrich) on AcLAPr activity were assessed by preincubating the enzyme with different concentrations of the reagents in 50 mM Tris-HCl buffer (pH 8.0) containing 1 mM MnCl2 for 20 min at 37°C, after which Leu-AMC (10 μM) was added and activity assayed as described above. .. The substrate specificity of AcLAPr was determined using Leu-AMC, l -arginine-AMC (Arg-AMC), and l -methionine-AMC (Met-AMC) (all from Sigma-Aldrich) as substrates.

    Infection:

    Article Title: The structure of avian polyomavirus reveals variably sized capsids, nonconserved inter-capsomere interactions, and a possible location of the minor capsid protein VP4
    Article Snippet: It was purified according to the protocol used by ; briefly, infected chicken embryonic cells were sonicated on ice in the presence of Freon-TF. .. L-arginine has been used to reduce protein aggregation , and we treated purified APV with GP buffer plus 250 mM L-arginine (APV+R) by dialysis (D-tube Mini Dialyzers, MWCO 12-14 kDa, Novagen) to stabilize and de-aggregate particles.

    Expressing:

    Article Title: HLA-B57 micropolymorphism defines the sequence and conformational breadth of the immunopeptidome
    Article Snippet: The HLA-B*57:01 , HLA-B*57:03 , HLA-B*58:01 and β 2 m genes were sub-cloned into the pET-30 expression vector and were expressed into inclusion bodies separately in Escherichia coli . .. Briefly, 90 mg HLA heavy chain was refolded by rapid dilution in a solution containing 3 M urea (Sigma-Aldrich, USA), 100 mM Tris-HCl, pH 8.0 (Sigma-Aldrich, USA), 400 mM l -arginine-HCl, 5 mM reduced glutathione (Sigma-Aldrich, USA) and 0.5 mM oxidised glutathione (Sigma-Aldrich, USA) in the presence of 30 mg β2 m and 10 mg of the appropriate peptide for 48 h. The refolded HLA-peptide complexes were dialysed into 10 mM Tris, pH 8.0, and purified by size-exclusion chromatography using HiLoad 16/60 Superdex 200 pg (GE Healthcare, USA) columns on an AKTA Purifier (GE Healthcare, USA) FPLC chromatography systems in 10 mM Tris, pH 8.0, and 150 mM NaCl buffer.

    Knock-In:

    Article Title: Assessing the impact of minimizing arginine conversion in fully defined SILAC culture medium in human embryonic stem cells
    Article Snippet: 2.2 Feeder‐free culture of hESC WA01 OCT4–eGFP Knock‐In hESC (WiCell Research Institute) were plated on a precoated xeno‐free vitronectin (VN XF, Primorigen Biosciences, Madison, WI, USA) 6‐well plate (coating concentration = 0.5 μg/cm2 ) and cultured in E8TM medium (37°C, 5% CO2 , and 5% O2 ). .. E8TM medium was made by diluting E8TM 50× supplement 1:50 with “arginine and lysine‐free” DMEM/F12 (Thermo Scientific, Rockford, IL, USA) supplemented with 398 μM l‐ arginine HCl and 499 μM l‐ lysine HCl (both from Sigma‐Aldrich, St. Louis, MO, USA).

    Flow Cytometry:

    Article Title: HLA-B57 micropolymorphism defines the sequence and conformational breadth of the immunopeptidome
    Article Snippet: Briefly, 90 mg HLA heavy chain was refolded by rapid dilution in a solution containing 3 M urea (Sigma-Aldrich, USA), 100 mM Tris-HCl, pH 8.0 (Sigma-Aldrich, USA), 400 mM l -arginine-HCl, 5 mM reduced glutathione (Sigma-Aldrich, USA) and 0.5 mM oxidised glutathione (Sigma-Aldrich, USA) in the presence of 30 mg β2 m and 10 mg of the appropriate peptide for 48 h. The refolded HLA-peptide complexes were dialysed into 10 mM Tris, pH 8.0, and purified by size-exclusion chromatography using HiLoad 16/60 Superdex 200 pg (GE Healthcare, USA) columns on an AKTA Purifier (GE Healthcare, USA) FPLC chromatography systems in 10 mM Tris, pH 8.0, and 150 mM NaCl buffer. .. Final purification was by anion exchange using a HiTrap Q Fast Flow column (GE Healthcare, USA) on the same AKTA system in 10 mM Tris pH 8.0 buffer with a NaCl gradient from 0 to 500 mM over 45 min.

    Chromatography:

    Article Title: HLA-B57 micropolymorphism defines the sequence and conformational breadth of the immunopeptidome
    Article Snippet: .. Briefly, 90 mg HLA heavy chain was refolded by rapid dilution in a solution containing 3 M urea (Sigma-Aldrich, USA), 100 mM Tris-HCl, pH 8.0 (Sigma-Aldrich, USA), 400 mM l -arginine-HCl, 5 mM reduced glutathione (Sigma-Aldrich, USA) and 0.5 mM oxidised glutathione (Sigma-Aldrich, USA) in the presence of 30 mg β2 m and 10 mg of the appropriate peptide for 48 h. The refolded HLA-peptide complexes were dialysed into 10 mM Tris, pH 8.0, and purified by size-exclusion chromatography using HiLoad 16/60 Superdex 200 pg (GE Healthcare, USA) columns on an AKTA Purifier (GE Healthcare, USA) FPLC chromatography systems in 10 mM Tris, pH 8.0, and 150 mM NaCl buffer. .. Final purification was by anion exchange using a HiTrap Q Fast Flow column (GE Healthcare, USA) on the same AKTA system in 10 mM Tris pH 8.0 buffer with a NaCl gradient from 0 to 500 mM over 45 min.

    Immunoprecipitation:

    Article Title: The enzymatic activity of 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase is enhanced by NPM-ALK: new insights in ALK-mediated pathogenesis and the treatment of ALCL
    Article Snippet: Cells were grown in RPMI medium lacking arginine and lysine, supplemented with 10% dialyzed fetal bovine serum, penicillin/streptomycin, and L-lysine/HCl and L-arginine/HCl (Sigma-Aldrich) for light cultures or L-arginine/HCL (U-13 C6 , 98%) and L-lysine/2HCl (U-13 C6 , 98%; Cambridge Isotope Laboratories, Andover, MA) for heavy cultures as described., Cells were grown to a density of approximately 106 cells/mL for a total of 108 cells per each cell culture type (2 × 108 cells total). .. After lysis, heavy and light cultures were combined and carried through the phosphopeptide immunoprecipitation protocol.

    Cell Culture:

    Article Title: The enzymatic activity of 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase is enhanced by NPM-ALK: new insights in ALK-mediated pathogenesis and the treatment of ALCL
    Article Snippet: .. Cells were grown in RPMI medium lacking arginine and lysine, supplemented with 10% dialyzed fetal bovine serum, penicillin/streptomycin, and L-lysine/HCl and L-arginine/HCl (Sigma-Aldrich) for light cultures or L-arginine/HCL (U-13 C6 , 98%) and L-lysine/2HCl (U-13 C6 , 98%; Cambridge Isotope Laboratories, Andover, MA) for heavy cultures as described., Cells were grown to a density of approximately 106 cells/mL for a total of 108 cells per each cell culture type (2 × 108 cells total). .. After lysis, heavy and light cultures were combined and carried through the phosphopeptide immunoprecipitation protocol.

    Article Title: Ets-1 is a transcriptional mediator of oncogenic nitric oxide signaling in estrogen receptor-negative breast cancer
    Article Snippet: Paragraph title: Cell culture and reagents ... Aminoguanidine (AG) and L-arginine (L-Arg) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Article Title: Assessing the impact of minimizing arginine conversion in fully defined SILAC culture medium in human embryonic stem cells
    Article Snippet: 2.2 Feeder‐free culture of hESC WA01 OCT4–eGFP Knock‐In hESC (WiCell Research Institute) were plated on a precoated xeno‐free vitronectin (VN XF, Primorigen Biosciences, Madison, WI, USA) 6‐well plate (coating concentration = 0.5 μg/cm2 ) and cultured in E8TM medium (37°C, 5% CO2 , and 5% O2 ). .. E8TM medium was made by diluting E8TM 50× supplement 1:50 with “arginine and lysine‐free” DMEM/F12 (Thermo Scientific, Rockford, IL, USA) supplemented with 398 μM l‐ arginine HCl and 499 μM l‐ lysine HCl (both from Sigma‐Aldrich, St. Louis, MO, USA).

    Article Title: Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function
    Article Snippet: .. Medium was supplemented with indicated concentrations of L-arginine (Sigma, #A5006) and/or L-ornithine (Sigma, #O2375) and cultured up to 72h before proceeding with intracellular cytokine staining. .. Flow cytometry To sample blood, 3-4 drops of blood were collected in 1 mL MEM medium (GIBCO) supplemented with 2000 U/L heparin (Heparin-Natrium-5000-ratiopharm).

    Article Title: Molecular Signature of CAID Syndrome: Noncanonical Roles of SGO1 in Regulation of TGF-β Signaling and Epigenomics
    Article Snippet: .. Cell culture and SILAC SILAC Dulbecco's modified Eagle medium without L-arginine and L-lysine (88364; Thermo Fisher Scientific, Waltham, MA) was supplemented with 13C6, 15N4 L-arginine (20104102; Silantes, München, Germany), 13C6, and 15N2 L-lysine (2111604102; Silantes), or unlabeled 12C614N4 L-arginine (A5006; Sigma Aldrich, St. Louis, MO) and 12C6L-lysine (L5501; Sigma Aldrich) to produce heavy and light SILAC media, respectively. .. L-proline (P5607; Sigma Aldrich) also was added to a final concentration of 100 μg/mL to prevent amino acid conversion.

    Article Title: Cell-specific Labeling Enzymes for Analysis of Cell–Cell Communication in Continuous Co-culture *
    Article Snippet: Paragraph title: Cell Culture ... All cells were obtained from ATCC Manassas, VA (except KPC cells, a kind gift from Professor Owen Sansom, Glasgow) and were grown in DMEM (deficient for L -lysine and L -arginine) (DMP49, Caisson, Utah) supplemented with 10% (v/v) dialyzed FBS (Invitrogen), 0.3 m m L -arginine (A8094, Sigma) and/or L -lysine (L5501, Sigma), D -lysine (L5876, Sigma), and DAP (07036, Sigma).

    Sonication:

    Article Title: The structure of avian polyomavirus reveals variably sized capsids, nonconserved inter-capsomere interactions, and a possible location of the minor capsid protein VP4
    Article Snippet: It was purified according to the protocol used by ; briefly, infected chicken embryonic cells were sonicated on ice in the presence of Freon-TF. .. L-arginine has been used to reduce protein aggregation , and we treated purified APV with GP buffer plus 250 mM L-arginine (APV+R) by dialysis (D-tube Mini Dialyzers, MWCO 12-14 kDa, Novagen) to stabilize and de-aggregate particles.

    Recombinant:

    Article Title: Ets-1 is a transcriptional mediator of oncogenic nitric oxide signaling in estrogen receptor-negative breast cancer
    Article Snippet: Aminoguanidine (AG) and L-arginine (L-Arg) were purchased from Sigma-Aldrich (St. Louis, MO, USA). .. Recombinant human epidermal growth factor (EGF) was purchased from R & D Systems (Minneapolis, MN, USA).

    Article Title: HLA-B57 micropolymorphism defines the sequence and conformational breadth of the immunopeptidome
    Article Snippet: Paragraph title: Recombinant HLA-peptide complex generation ... Briefly, 90 mg HLA heavy chain was refolded by rapid dilution in a solution containing 3 M urea (Sigma-Aldrich, USA), 100 mM Tris-HCl, pH 8.0 (Sigma-Aldrich, USA), 400 mM l -arginine-HCl, 5 mM reduced glutathione (Sigma-Aldrich, USA) and 0.5 mM oxidised glutathione (Sigma-Aldrich, USA) in the presence of 30 mg β2 m and 10 mg of the appropriate peptide for 48 h. The refolded HLA-peptide complexes were dialysed into 10 mM Tris, pH 8.0, and purified by size-exclusion chromatography using HiLoad 16/60 Superdex 200 pg (GE Healthcare, USA) columns on an AKTA Purifier (GE Healthcare, USA) FPLC chromatography systems in 10 mM Tris, pH 8.0, and 150 mM NaCl buffer.

    Molecular Weight:

    Article Title: The structure of avian polyomavirus reveals variably sized capsids, nonconserved inter-capsomere interactions, and a possible location of the minor capsid protein VP4
    Article Snippet: Viral bands were collected after spinning at 53,000 · g for 3 hours, and CsCl was removed by dialysis (D-tube Mini Dialyzers, MWCO (molecular weight cutoff) 12-14 kDa, Novagen, Gibbstown, New Jersey, USA) and the sample was retained in GP buffer (200 mM NaCl, 20 mM Tris-HCl, 1 mM CaCl2 buffer, pH 7.4). .. L-arginine has been used to reduce protein aggregation , and we treated purified APV with GP buffer plus 250 mM L-arginine (APV+R) by dialysis (D-tube Mini Dialyzers, MWCO 12-14 kDa, Novagen) to stabilize and de-aggregate particles.

    Fluorescence:

    Article Title: Essential Role for an M17 Leucine Aminopeptidase in Encystation of Acanthamoeba castellanii
    Article Snippet: Briefly, 10 μL of enzyme solution was added to 190 μL of assay buffer (50 mM Tris-HCl, 1 mM MnCl2 , pH 8.0) containing 10 μM Leu-AMC and incubated for 20 min at 37°C, and the release of fluorescence was measured at an excitation wavelength of 370 nm and an emission wavelength of 440 nm using a Gemini EM fluorescence microplate reader (Molecular Devices, Sunnyvale, CA, USA). .. The substrate specificity of AcLAPr was determined using Leu-AMC, l -arginine-AMC (Arg-AMC), and l -methionine-AMC (Met-AMC) (all from Sigma-Aldrich) as substrates.

    Multiple Displacement Amplification:

    Article Title: Ets-1 is a transcriptional mediator of oncogenic nitric oxide signaling in estrogen receptor-negative breast cancer
    Article Snippet: Cell culture and reagents Human breast adenocarcinoma cell lines MDA-MB-231, MDA-MB-468 and SKBR3 (American Type Culture Collection (ATCC), Manassas, VA, USA) were cultured in RPMI medium (Invitrogen, Carlsbad, CA, USA) containing 10% fetal bovine serum (Atlanta Biologics, Norcross, GA, USA) and 100 IU penicillin and 100 µg/ml streptomycin (Invitrogen). .. Aminoguanidine (AG) and L-arginine (L-Arg) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Isolation:

    Article Title: Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function
    Article Snippet: Cells were counted and CD8 T cells isolated using a magnetic activated cell sorting negative selection (MACS) kit according to the manufacturer’s instructions (Miltenyi Biotec, #130-104-075). .. Medium was supplemented with indicated concentrations of L-arginine (Sigma, #A5006) and/or L-ornithine (Sigma, #O2375) and cultured up to 72h before proceeding with intracellular cytokine staining.

    Size-exclusion Chromatography:

    Article Title: HLA-B57 micropolymorphism defines the sequence and conformational breadth of the immunopeptidome
    Article Snippet: .. Briefly, 90 mg HLA heavy chain was refolded by rapid dilution in a solution containing 3 M urea (Sigma-Aldrich, USA), 100 mM Tris-HCl, pH 8.0 (Sigma-Aldrich, USA), 400 mM l -arginine-HCl, 5 mM reduced glutathione (Sigma-Aldrich, USA) and 0.5 mM oxidised glutathione (Sigma-Aldrich, USA) in the presence of 30 mg β2 m and 10 mg of the appropriate peptide for 48 h. The refolded HLA-peptide complexes were dialysed into 10 mM Tris, pH 8.0, and purified by size-exclusion chromatography using HiLoad 16/60 Superdex 200 pg (GE Healthcare, USA) columns on an AKTA Purifier (GE Healthcare, USA) FPLC chromatography systems in 10 mM Tris, pH 8.0, and 150 mM NaCl buffer. .. Final purification was by anion exchange using a HiTrap Q Fast Flow column (GE Healthcare, USA) on the same AKTA system in 10 mM Tris pH 8.0 buffer with a NaCl gradient from 0 to 500 mM over 45 min.

    Article Title: Insert engineering and solubility screening improves recovery of virus-like particle subunits displaying hydrophobic epitopes
    Article Snippet: .. Modified DD IT1 capsomere recovered via SEC in buffer containing 50 m M l -arginine ( l -Arg) (≥98%, Sigma-Aldrich, ) at pH 8.5 was assembled by dialysis in standard assembly buffer containing 0.5 M (NH4 )2 SO4 , 20 m M Tris–base, 5% glycerol (v/v), 1 m M CaCl2 at pH 7.4, with or without l -Arg at 50 m M . .. Samples were placed on 200-mesh carbon-coated copper grids and left to settle for 90 s. Following two washes in water, grids were negative stained with 2% uranyl acetate for 60 s. TEM was carried out on a JEOL 1010 at 80 kV and images were cropped and adjusted for brightness using Adobe Photoshop (CS6; ).

    Labeling:

    Article Title: The enzymatic activity of 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase is enhanced by NPM-ALK: new insights in ALK-mediated pathogenesis and the treatment of ALCL
    Article Snippet: Paragraph title: Stable isotope labeling of amino acid in cell culture analysis of shALK cells ... Cells were grown in RPMI medium lacking arginine and lysine, supplemented with 10% dialyzed fetal bovine serum, penicillin/streptomycin, and L-lysine/HCl and L-arginine/HCl (Sigma-Aldrich) for light cultures or L-arginine/HCL (U-13 C6 , 98%) and L-lysine/2HCl (U-13 C6 , 98%; Cambridge Isotope Laboratories, Andover, MA) for heavy cultures as described., Cells were grown to a density of approximately 106 cells/mL for a total of 108 cells per each cell culture type (2 × 108 cells total).

    Purification:

    Article Title: The structure of avian polyomavirus reveals variably sized capsids, nonconserved inter-capsomere interactions, and a possible location of the minor capsid protein VP4
    Article Snippet: .. L-arginine has been used to reduce protein aggregation , and we treated purified APV with GP buffer plus 250 mM L-arginine (APV+R) by dialysis (D-tube Mini Dialyzers, MWCO 12-14 kDa, Novagen) to stabilize and de-aggregate particles. .. The sample was subjected to centrifugal filtration (YM-100 centrifugal devices, Millipore, Billerica, Massachusetts, USA) for a final washing and concentrating step.

    Article Title: Biochemical enrichment and biophysical characterization of a taste receptor for L-arginine from the catfish, Ictalurus puntatus
    Article Snippet: Water was deionized and further purified through a Milli-Q Plus PF system (Bedford, MA). .. The absolute enantiomer, L-arginine HCl, was purchased from Sigma, and the absolute enantiomer, D-arginine HCl, was a gift of the Ajinomoto Co., Tokyo, Japan.

    Article Title: HLA-B57 micropolymorphism defines the sequence and conformational breadth of the immunopeptidome
    Article Snippet: .. Briefly, 90 mg HLA heavy chain was refolded by rapid dilution in a solution containing 3 M urea (Sigma-Aldrich, USA), 100 mM Tris-HCl, pH 8.0 (Sigma-Aldrich, USA), 400 mM l -arginine-HCl, 5 mM reduced glutathione (Sigma-Aldrich, USA) and 0.5 mM oxidised glutathione (Sigma-Aldrich, USA) in the presence of 30 mg β2 m and 10 mg of the appropriate peptide for 48 h. The refolded HLA-peptide complexes were dialysed into 10 mM Tris, pH 8.0, and purified by size-exclusion chromatography using HiLoad 16/60 Superdex 200 pg (GE Healthcare, USA) columns on an AKTA Purifier (GE Healthcare, USA) FPLC chromatography systems in 10 mM Tris, pH 8.0, and 150 mM NaCl buffer. .. Final purification was by anion exchange using a HiTrap Q Fast Flow column (GE Healthcare, USA) on the same AKTA system in 10 mM Tris pH 8.0 buffer with a NaCl gradient from 0 to 500 mM over 45 min.

    Positron Emission Tomography:

    Article Title: HLA-B57 micropolymorphism defines the sequence and conformational breadth of the immunopeptidome
    Article Snippet: The HLA-B*57:01 , HLA-B*57:03 , HLA-B*58:01 and β 2 m genes were sub-cloned into the pET-30 expression vector and were expressed into inclusion bodies separately in Escherichia coli . .. Briefly, 90 mg HLA heavy chain was refolded by rapid dilution in a solution containing 3 M urea (Sigma-Aldrich, USA), 100 mM Tris-HCl, pH 8.0 (Sigma-Aldrich, USA), 400 mM l -arginine-HCl, 5 mM reduced glutathione (Sigma-Aldrich, USA) and 0.5 mM oxidised glutathione (Sigma-Aldrich, USA) in the presence of 30 mg β2 m and 10 mg of the appropriate peptide for 48 h. The refolded HLA-peptide complexes were dialysed into 10 mM Tris, pH 8.0, and purified by size-exclusion chromatography using HiLoad 16/60 Superdex 200 pg (GE Healthcare, USA) columns on an AKTA Purifier (GE Healthcare, USA) FPLC chromatography systems in 10 mM Tris, pH 8.0, and 150 mM NaCl buffer.

    Fast Protein Liquid Chromatography:

    Article Title: HLA-B57 micropolymorphism defines the sequence and conformational breadth of the immunopeptidome
    Article Snippet: .. Briefly, 90 mg HLA heavy chain was refolded by rapid dilution in a solution containing 3 M urea (Sigma-Aldrich, USA), 100 mM Tris-HCl, pH 8.0 (Sigma-Aldrich, USA), 400 mM l -arginine-HCl, 5 mM reduced glutathione (Sigma-Aldrich, USA) and 0.5 mM oxidised glutathione (Sigma-Aldrich, USA) in the presence of 30 mg β2 m and 10 mg of the appropriate peptide for 48 h. The refolded HLA-peptide complexes were dialysed into 10 mM Tris, pH 8.0, and purified by size-exclusion chromatography using HiLoad 16/60 Superdex 200 pg (GE Healthcare, USA) columns on an AKTA Purifier (GE Healthcare, USA) FPLC chromatography systems in 10 mM Tris, pH 8.0, and 150 mM NaCl buffer. .. Final purification was by anion exchange using a HiTrap Q Fast Flow column (GE Healthcare, USA) on the same AKTA system in 10 mM Tris pH 8.0 buffer with a NaCl gradient from 0 to 500 mM over 45 min.

    Staining:

    Article Title: Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function
    Article Snippet: .. Medium was supplemented with indicated concentrations of L-arginine (Sigma, #A5006) and/or L-ornithine (Sigma, #O2375) and cultured up to 72h before proceeding with intracellular cytokine staining. .. Flow cytometry To sample blood, 3-4 drops of blood were collected in 1 mL MEM medium (GIBCO) supplemented with 2000 U/L heparin (Heparin-Natrium-5000-ratiopharm).

    Mouse Assay:

    Article Title: Disruption of Small GTPase Rab7 Exacerbates the Severity of Acute Pancreatitis in Experimental Mouse Models
    Article Snippet: Caerulein- and L-arginine-induced pancreatitis Acute pancreatitis was induced in age-matched 8- to 12-week-old male mice using caerulein or L-arginine. .. L-arginine-induced acute pancreatitis was triggered by injecting 4 g/kg (body weight) L-arginine (Sigma-Aldrich) intraperitoneally twice hourly.

    SDS Page:

    Article Title: The structure of avian polyomavirus reveals variably sized capsids, nonconserved inter-capsomere interactions, and a possible location of the minor capsid protein VP4
    Article Snippet: L-arginine has been used to reduce protein aggregation , and we treated purified APV with GP buffer plus 250 mM L-arginine (APV+R) by dialysis (D-tube Mini Dialyzers, MWCO 12-14 kDa, Novagen) to stabilize and de-aggregate particles. .. Purified APV was subjected to SDS-PAGE and immunoblotting as previously described ( ; ).

    Plasmid Preparation:

    Article Title: Biochemical enrichment and biophysical characterization of a taste receptor for L-arginine from the catfish, Ictalurus puntatus
    Article Snippet: The absolute enantiomer, L-arginine HCl, was purchased from Sigma, and the absolute enantiomer, D-arginine HCl, was a gift of the Ajinomoto Co., Tokyo, Japan. .. The absolute enantiomer, L-arginine HCl, was purchased from Sigma, and the absolute enantiomer, D-arginine HCl, was a gift of the Ajinomoto Co., Tokyo, Japan.

    Article Title: HLA-B57 micropolymorphism defines the sequence and conformational breadth of the immunopeptidome
    Article Snippet: The HLA-B*57:01 , HLA-B*57:03 , HLA-B*58:01 and β 2 m genes were sub-cloned into the pET-30 expression vector and were expressed into inclusion bodies separately in Escherichia coli . .. Briefly, 90 mg HLA heavy chain was refolded by rapid dilution in a solution containing 3 M urea (Sigma-Aldrich, USA), 100 mM Tris-HCl, pH 8.0 (Sigma-Aldrich, USA), 400 mM l -arginine-HCl, 5 mM reduced glutathione (Sigma-Aldrich, USA) and 0.5 mM oxidised glutathione (Sigma-Aldrich, USA) in the presence of 30 mg β2 m and 10 mg of the appropriate peptide for 48 h. The refolded HLA-peptide complexes were dialysed into 10 mM Tris, pH 8.0, and purified by size-exclusion chromatography using HiLoad 16/60 Superdex 200 pg (GE Healthcare, USA) columns on an AKTA Purifier (GE Healthcare, USA) FPLC chromatography systems in 10 mM Tris, pH 8.0, and 150 mM NaCl buffer.

    Selection:

    Article Title: Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function
    Article Snippet: Cells were counted and CD8 T cells isolated using a magnetic activated cell sorting negative selection (MACS) kit according to the manufacturer’s instructions (Miltenyi Biotec, #130-104-075). .. Medium was supplemented with indicated concentrations of L-arginine (Sigma, #A5006) and/or L-ornithine (Sigma, #O2375) and cultured up to 72h before proceeding with intracellular cytokine staining.

    In Vitro:

    Article Title: Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function
    Article Snippet: Paragraph title: CD8 T cell isolation and in vitro activation ... Medium was supplemented with indicated concentrations of L-arginine (Sigma, #A5006) and/or L-ornithine (Sigma, #O2375) and cultured up to 72h before proceeding with intracellular cytokine staining.

    Article Title: Insert engineering and solubility screening improves recovery of virus-like particle subunits displaying hydrophobic epitopes
    Article Snippet: Paragraph title: In vitro VLP assembly ... Modified DD IT1 capsomere recovered via SEC in buffer containing 50 m M l -arginine ( l -Arg) (≥98%, Sigma-Aldrich, ) at pH 8.5 was assembled by dialysis in standard assembly buffer containing 0.5 M (NH4 )2 SO4 , 20 m M Tris–base, 5% glycerol (v/v), 1 m M CaCl2 at pH 7.4, with or without l -Arg at 50 m M .

    Activation Assay:

    Article Title: Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function
    Article Snippet: Paragraph title: CD8 T cell isolation and in vitro activation ... Medium was supplemented with indicated concentrations of L-arginine (Sigma, #A5006) and/or L-ornithine (Sigma, #O2375) and cultured up to 72h before proceeding with intracellular cytokine staining.

    Concentration Assay:

    Article Title: Assessing the impact of minimizing arginine conversion in fully defined SILAC culture medium in human embryonic stem cells
    Article Snippet: 2.2 Feeder‐free culture of hESC WA01 OCT4–eGFP Knock‐In hESC (WiCell Research Institute) were plated on a precoated xeno‐free vitronectin (VN XF, Primorigen Biosciences, Madison, WI, USA) 6‐well plate (coating concentration = 0.5 μg/cm2 ) and cultured in E8TM medium (37°C, 5% CO2 , and 5% O2 ). .. E8TM medium was made by diluting E8TM 50× supplement 1:50 with “arginine and lysine‐free” DMEM/F12 (Thermo Scientific, Rockford, IL, USA) supplemented with 398 μM l‐ arginine HCl and 499 μM l‐ lysine HCl (both from Sigma‐Aldrich, St. Louis, MO, USA).

    Article Title: Molecular Signature of CAID Syndrome: Noncanonical Roles of SGO1 in Regulation of TGF-β Signaling and Epigenomics
    Article Snippet: Cell culture and SILAC SILAC Dulbecco's modified Eagle medium without L-arginine and L-lysine (88364; Thermo Fisher Scientific, Waltham, MA) was supplemented with 13C6, 15N4 L-arginine (20104102; Silantes, München, Germany), 13C6, and 15N2 L-lysine (2111604102; Silantes), or unlabeled 12C614N4 L-arginine (A5006; Sigma Aldrich, St. Louis, MO) and 12C6L-lysine (L5501; Sigma Aldrich) to produce heavy and light SILAC media, respectively. .. L-proline (P5607; Sigma Aldrich) also was added to a final concentration of 100 μg/mL to prevent amino acid conversion.

    Magnetic Cell Separation:

    Article Title: Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function
    Article Snippet: Cells were counted and CD8 T cells isolated using a magnetic activated cell sorting negative selection (MACS) kit according to the manufacturer’s instructions (Miltenyi Biotec, #130-104-075). .. Medium was supplemented with indicated concentrations of L-arginine (Sigma, #A5006) and/or L-ornithine (Sigma, #O2375) and cultured up to 72h before proceeding with intracellular cytokine staining.

    Lysis:

    Article Title: The enzymatic activity of 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase is enhanced by NPM-ALK: new insights in ALK-mediated pathogenesis and the treatment of ALCL
    Article Snippet: Cells were grown in RPMI medium lacking arginine and lysine, supplemented with 10% dialyzed fetal bovine serum, penicillin/streptomycin, and L-lysine/HCl and L-arginine/HCl (Sigma-Aldrich) for light cultures or L-arginine/HCL (U-13 C6 , 98%) and L-lysine/2HCl (U-13 C6 , 98%; Cambridge Isotope Laboratories, Andover, MA) for heavy cultures as described., Cells were grown to a density of approximately 106 cells/mL for a total of 108 cells per each cell culture type (2 × 108 cells total). .. After lysis, heavy and light cultures were combined and carried through the phosphopeptide immunoprecipitation protocol.

    Article Title: Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function
    Article Snippet: The pellet was resuspended in 1 mL red blood cell lysis buffer (eBioscience, #00-4300-54) and incubated at room temperature for 1 min. .. Medium was supplemented with indicated concentrations of L-arginine (Sigma, #A5006) and/or L-ornithine (Sigma, #O2375) and cultured up to 72h before proceeding with intracellular cytokine staining.

    FACS:

    Article Title: Type I Interferon Signaling Disrupts the Hepatic Urea Cycle and Alters Systemic Metabolism to Suppress T Cell Function
    Article Snippet: Cells were counted and CD8 T cells isolated using a magnetic activated cell sorting negative selection (MACS) kit according to the manufacturer’s instructions (Miltenyi Biotec, #130-104-075). .. Medium was supplemented with indicated concentrations of L-arginine (Sigma, #A5006) and/or L-ornithine (Sigma, #O2375) and cultured up to 72h before proceeding with intracellular cytokine staining.

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    Millipore enos inhibitor ng nitro l arginine methyl ester l name
    P1 failed to enhance <t>eNOS</t> activity and angiogenesis in the presence of eNOS inhibitor <t>L-NAME</t> or PKC-α inhibitor GO6979 (GO). ECs were pretreated overnight with L-NAME (1 mM) or GO (10 μM) in RPMI 1640 followed by 60 min at 37°C
    Enos Inhibitor Ng Nitro L Arginine Methyl Ester L Name, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/enos inhibitor ng nitro l arginine methyl ester l name/product/Millipore
    Average 88 stars, based on 1 article reviews
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    enos inhibitor ng nitro l arginine methyl ester l name - by Bioz Stars, 2020-04
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    86
    Millipore n g nitro d arginine
    Endocannabinoid-stimulated NO accumulation antagonized by a NOS inhibitor. a N18TG2 cells were treated with a methanandamide (MetAEA) alone (1 μM) or b following pretreatment with NOS inhibitor <t>N</t> G -nitro- l -arginine ( l- NNA ; 1 μM; bar = 100 μm). b Quantitation of effects of NOS inhibitor l- NNA and its inactive enantiomer N G -nitro- d -arginine ( d -NNA ; 1 μM) on methanandamide-stimulated NO accumulation in N18TG2 cells. Data are presented as the DAF/DAPI fluorescence intensity ratio (mean ± SEM from N = 3 independent experiments). Values significantly different from those of MetAEA-stimulated cells (* p
    N G Nitro D Arginine, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n g nitro d arginine/product/Millipore
    Average 86 stars, based on 3 article reviews
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    n g nitro d arginine - by Bioz Stars, 2020-04
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    94
    Millipore n g nitro l arginine methyl ester
    Group means ± SE of vasodilation response [percent of maximal vasodilation (%CVC max )] to increased core temperature (ΔT or ) at baseline (T or = 0.0) and during whole body heating in Ringer's control ( A ), BH 4 -perfused ( B ), and <t>N</t> G -nitro- l
    N G Nitro L Arginine Methyl Ester, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n g nitro l arginine methyl ester/product/Millipore
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    n g nitro l arginine methyl ester - by Bioz Stars, 2020-04
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    Image Search Results


    P1 failed to enhance eNOS activity and angiogenesis in the presence of eNOS inhibitor L-NAME or PKC-α inhibitor GO6979 (GO). ECs were pretreated overnight with L-NAME (1 mM) or GO (10 μM) in RPMI 1640 followed by 60 min at 37°C

    Journal: Nitric oxide : biology and chemistry / official journal of the Nitric Oxide Society

    Article Title: Peptide-Stimulated Angiogenesis: Role of Lung Endothelial Caveolar Signaling and Nitric Oxide

    doi: 10.1016/j.niox.2015.10.002

    Figure Lengend Snippet: P1 failed to enhance eNOS activity and angiogenesis in the presence of eNOS inhibitor L-NAME or PKC-α inhibitor GO6979 (GO). ECs were pretreated overnight with L-NAME (1 mM) or GO (10 μM) in RPMI 1640 followed by 60 min at 37°C

    Article Snippet: PI3K inhibitor Wortmannin (WT), eNOS inhibitor NG -nitro-L-arginine methyl ester (L-NAME), PKC-α inhibitor GO6976, and NO donor NOC-18 were obtained from Calbiochem (Gibbstown.

    Techniques: Activity Assay

    Endocannabinoid-stimulated NO accumulation antagonized by a NOS inhibitor. a N18TG2 cells were treated with a methanandamide (MetAEA) alone (1 μM) or b following pretreatment with NOS inhibitor N G -nitro- l -arginine ( l- NNA ; 1 μM; bar = 100 μm). b Quantitation of effects of NOS inhibitor l- NNA and its inactive enantiomer N G -nitro- d -arginine ( d -NNA ; 1 μM) on methanandamide-stimulated NO accumulation in N18TG2 cells. Data are presented as the DAF/DAPI fluorescence intensity ratio (mean ± SEM from N = 3 independent experiments). Values significantly different from those of MetAEA-stimulated cells (* p

    Journal: Journal of Neuroimmune Pharmacology

    Article Title: Cannabinoid Regulation of Nitric Oxide Synthase I (nNOS) in Neuronal Cells

    doi: 10.1007/s11481-009-9153-7

    Figure Lengend Snippet: Endocannabinoid-stimulated NO accumulation antagonized by a NOS inhibitor. a N18TG2 cells were treated with a methanandamide (MetAEA) alone (1 μM) or b following pretreatment with NOS inhibitor N G -nitro- l -arginine ( l- NNA ; 1 μM; bar = 100 μm). b Quantitation of effects of NOS inhibitor l- NNA and its inactive enantiomer N G -nitro- d -arginine ( d -NNA ; 1 μM) on methanandamide-stimulated NO accumulation in N18TG2 cells. Data are presented as the DAF/DAPI fluorescence intensity ratio (mean ± SEM from N = 3 independent experiments). Values significantly different from those of MetAEA-stimulated cells (* p

    Article Snippet: N G -nitro-l -arginine (l- NNA) and N G -nitro-d -arginine (d -NNA) were purchased from EMD Chemicals (Gibbstown, NJ, USA).

    Techniques: Quantitation Assay, Fluorescence

    Group means ± SE of vasodilation response [percent of maximal vasodilation (%CVC max )] to increased core temperature (ΔT or ) at baseline (T or = 0.0) and during whole body heating in Ringer's control ( A ), BH 4 -perfused ( B ), and N G -nitro- l

    Journal: Journal of Applied Physiology

    Article Title: Oral sapropterin acutely augments reflex vasodilation in aged human skin through nitric oxide-dependent mechanisms

    doi: 10.1152/japplphysiol.00481.2013

    Figure Lengend Snippet: Group means ± SE of vasodilation response [percent of maximal vasodilation (%CVC max )] to increased core temperature (ΔT or ) at baseline (T or = 0.0) and during whole body heating in Ringer's control ( A ), BH 4 -perfused ( B ), and N G -nitro- l

    Article Snippet: The MD fibers were randomly assigned to deliver 1 ) lactated Ringer's solution to serve as control, 2 ) 10 mM BH4 (Sigma, St. Louis, MO) for local BH4 administration, or 3 ) 20 mM N G -nitro- l -arginine methyl ester ( l -NAME; Calbiochem, San Diego, CA) to inhibit NOS.

    Techniques: