nω nitro l arginine  (Millipore)


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    Structured Review

    Millipore nω nitro l arginine
    Exercise training-induced remodeling of rat intramural coronary arterioles compared with sedentary controls. a Inner diameter in spontaneous tone and myogenic constriction expressed as a ratio of the passive diameter. The dashed line refers to the diameter of arterioles in the passive condition, taken as the reference. Alteration of the spontaneously contracted inner diameter in response to the NOS inhibitor N ω <t>-nitro-L-arginine</t> (L-NNA, 10 −5 M; b ) and the cyclo-oxygenase inhibitor (INDO, 2.8 × 10 −5 M; c ). The dotted line ( b , c ), representing the diameter of arterioles in control myogenic constriction, was taken as a reference at each pressure level. Mean values were used from 2-way ANOVA tests with Tukey paired comparisons: * p
    Nω Nitro L Arginine, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 8 article reviews
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    nω nitro l arginine - by Bioz Stars, 2020-04
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    Images

    1) Product Images from "Remodeling of Wall Mechanics and the Myogenic Mechanism of Rat Intramural Coronary Arterioles in Response to a Short-Term Daily Exercise Program: Role of Endothelial Factors"

    Article Title: Remodeling of Wall Mechanics and the Myogenic Mechanism of Rat Intramural Coronary Arterioles in Response to a Short-Term Daily Exercise Program: Role of Endothelial Factors

    Journal: Journal of vascular research

    doi: 10.1159/000486571

    Exercise training-induced remodeling of rat intramural coronary arterioles compared with sedentary controls. a Inner diameter in spontaneous tone and myogenic constriction expressed as a ratio of the passive diameter. The dashed line refers to the diameter of arterioles in the passive condition, taken as the reference. Alteration of the spontaneously contracted inner diameter in response to the NOS inhibitor N ω -nitro-L-arginine (L-NNA, 10 −5 M; b ) and the cyclo-oxygenase inhibitor (INDO, 2.8 × 10 −5 M; c ). The dotted line ( b , c ), representing the diameter of arterioles in control myogenic constriction, was taken as a reference at each pressure level. Mean values were used from 2-way ANOVA tests with Tukey paired comparisons: * p
    Figure Legend Snippet: Exercise training-induced remodeling of rat intramural coronary arterioles compared with sedentary controls. a Inner diameter in spontaneous tone and myogenic constriction expressed as a ratio of the passive diameter. The dashed line refers to the diameter of arterioles in the passive condition, taken as the reference. Alteration of the spontaneously contracted inner diameter in response to the NOS inhibitor N ω -nitro-L-arginine (L-NNA, 10 −5 M; b ) and the cyclo-oxygenase inhibitor (INDO, 2.8 × 10 −5 M; c ). The dotted line ( b , c ), representing the diameter of arterioles in control myogenic constriction, was taken as a reference at each pressure level. Mean values were used from 2-way ANOVA tests with Tukey paired comparisons: * p

    Techniques Used:

    Exercise training-induced remodeling of rat intramural coronary arterioles. The inner diameter as a function of increasing transmural pressure was raised in a stepwise manner. Spontaneous tone and myogenic constriction in the control condition (green, trained; yellow, sedentary; a , b ), in the effect of the NOS inhibitor N ω -nitro-L-arginine (L-NNA, 10 −5 M; blue, trained; magenta, sedentary; a ), and in the effect of the cyclo-oxygenase inhibitor indomethacin (INDO, 2.8 × 10 −5 M; b ). Passive pressure-diameter curves are also shown for easy comparison (black, trained; red, sedentary; a , b ). Mean values were used from 2-way ANOVA tests with Tukey paired comparisons: * p
    Figure Legend Snippet: Exercise training-induced remodeling of rat intramural coronary arterioles. The inner diameter as a function of increasing transmural pressure was raised in a stepwise manner. Spontaneous tone and myogenic constriction in the control condition (green, trained; yellow, sedentary; a , b ), in the effect of the NOS inhibitor N ω -nitro-L-arginine (L-NNA, 10 −5 M; blue, trained; magenta, sedentary; a ), and in the effect of the cyclo-oxygenase inhibitor indomethacin (INDO, 2.8 × 10 −5 M; b ). Passive pressure-diameter curves are also shown for easy comparison (black, trained; red, sedentary; a , b ). Mean values were used from 2-way ANOVA tests with Tukey paired comparisons: * p

    Techniques Used:

    Exercise training-induced remodeling of rat intramural coronary arterioles: a schematic representation. Wall sections of exercised and sedentary male rat coronary arteriole segments in spontaneous myogenic tone without ( a ) and with ( b , c ) inhibitors. a Coronary arterioles in control myogenic tone at 120 mm Hg intraluminal pressure and with the passive diameter. b The effect of the NO inhibitor N ω -nitro-L-arginine (in the presence of prostanoids) reveals an increased (NO-mediated) dilation effect in myogenic tone at 30 mm Hg intraluminal pressure in the coronary arterioles of exercised rats. c The effect of the cyclo-oxygenase inhibitor INDO (in the presence of NO) reveals the presence of endogenous constrictor prostanoids at 120 mm Hg intraluminal pressure in the coronary arterioles of sedentary animals, which is missing in the exercised group.
    Figure Legend Snippet: Exercise training-induced remodeling of rat intramural coronary arterioles: a schematic representation. Wall sections of exercised and sedentary male rat coronary arteriole segments in spontaneous myogenic tone without ( a ) and with ( b , c ) inhibitors. a Coronary arterioles in control myogenic tone at 120 mm Hg intraluminal pressure and with the passive diameter. b The effect of the NO inhibitor N ω -nitro-L-arginine (in the presence of prostanoids) reveals an increased (NO-mediated) dilation effect in myogenic tone at 30 mm Hg intraluminal pressure in the coronary arterioles of exercised rats. c The effect of the cyclo-oxygenase inhibitor INDO (in the presence of NO) reveals the presence of endogenous constrictor prostanoids at 120 mm Hg intraluminal pressure in the coronary arterioles of sedentary animals, which is missing in the exercised group.

    Techniques Used:

    2) Product Images from "Hydrogen Peroxide and Nitric Oxide are Involved in Salicylic Acid-Induced Salvianolic Acid B Production in Salvia miltiorrhiza Cell Cultures"

    Article Title: Hydrogen Peroxide and Nitric Oxide are Involved in Salicylic Acid-Induced Salvianolic Acid B Production in Salvia miltiorrhiza Cell Cultures

    Journal: Molecules

    doi: 10.3390/molecules19055913

    Effects of SA, sodium nitroprusside (SNP), N ω -nitro-L-arginine (L-NNA), carboxy-2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) and complex treatments on the production of Sal B and NO in Salvia miltiorrhiza cell cultures. The concentrations for SA, SNP, L-NNA and cPTIO were 22 mg L −1 , 0.5 mmol L −1 , 2 mmol L −1 and 50 μmol L −1 , respectively. SA was added to subcultured Salvia miltiorrhiza cells after 8-d culture, and in the subsequent 4 h and 2 d, both contents of NO and Sal B were determined, respectively, with three replications. Different lowercase letters represented significance at 0.05.
    Figure Legend Snippet: Effects of SA, sodium nitroprusside (SNP), N ω -nitro-L-arginine (L-NNA), carboxy-2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) and complex treatments on the production of Sal B and NO in Salvia miltiorrhiza cell cultures. The concentrations for SA, SNP, L-NNA and cPTIO were 22 mg L −1 , 0.5 mmol L −1 , 2 mmol L −1 and 50 μmol L −1 , respectively. SA was added to subcultured Salvia miltiorrhiza cells after 8-d culture, and in the subsequent 4 h and 2 d, both contents of NO and Sal B were determined, respectively, with three replications. Different lowercase letters represented significance at 0.05.

    Techniques Used:

    3) Product Images from "Remodeling of Wall Mechanics and the Myogenic Mechanism of Rat Intramural Coronary Arterioles in Response to a Short-Term Daily Exercise Program: Role of Endothelial Factors"

    Article Title: Remodeling of Wall Mechanics and the Myogenic Mechanism of Rat Intramural Coronary Arterioles in Response to a Short-Term Daily Exercise Program: Role of Endothelial Factors

    Journal: Journal of vascular research

    doi: 10.1159/000486571

    Exercise training-induced remodeling of rat intramural coronary arterioles compared with sedentary controls. a Inner diameter in spontaneous tone and myogenic constriction expressed as a ratio of the passive diameter. The dashed line refers to the diameter of arterioles in the passive condition, taken as the reference. Alteration of the spontaneously contracted inner diameter in response to the NOS inhibitor N ω -nitro-L-arginine (L-NNA, 10 −5 M; b ) and the cyclo-oxygenase inhibitor (INDO, 2.8 × 10 −5 M; c ). The dotted line ( b , c ), representing the diameter of arterioles in control myogenic constriction, was taken as a reference at each pressure level. Mean values were used from 2-way ANOVA tests with Tukey paired comparisons: * p
    Figure Legend Snippet: Exercise training-induced remodeling of rat intramural coronary arterioles compared with sedentary controls. a Inner diameter in spontaneous tone and myogenic constriction expressed as a ratio of the passive diameter. The dashed line refers to the diameter of arterioles in the passive condition, taken as the reference. Alteration of the spontaneously contracted inner diameter in response to the NOS inhibitor N ω -nitro-L-arginine (L-NNA, 10 −5 M; b ) and the cyclo-oxygenase inhibitor (INDO, 2.8 × 10 −5 M; c ). The dotted line ( b , c ), representing the diameter of arterioles in control myogenic constriction, was taken as a reference at each pressure level. Mean values were used from 2-way ANOVA tests with Tukey paired comparisons: * p

    Techniques Used:

    Exercise training-induced remodeling of rat intramural coronary arterioles. The inner diameter as a function of increasing transmural pressure was raised in a stepwise manner. Spontaneous tone and myogenic constriction in the control condition (green, trained; yellow, sedentary; a , b ), in the effect of the NOS inhibitor N ω -nitro-L-arginine (L-NNA, 10 −5 M; blue, trained; magenta, sedentary; a ), and in the effect of the cyclo-oxygenase inhibitor indomethacin (INDO, 2.8 × 10 −5 M; b ). Passive pressure-diameter curves are also shown for easy comparison (black, trained; red, sedentary; a , b ). Mean values were used from 2-way ANOVA tests with Tukey paired comparisons: * p
    Figure Legend Snippet: Exercise training-induced remodeling of rat intramural coronary arterioles. The inner diameter as a function of increasing transmural pressure was raised in a stepwise manner. Spontaneous tone and myogenic constriction in the control condition (green, trained; yellow, sedentary; a , b ), in the effect of the NOS inhibitor N ω -nitro-L-arginine (L-NNA, 10 −5 M; blue, trained; magenta, sedentary; a ), and in the effect of the cyclo-oxygenase inhibitor indomethacin (INDO, 2.8 × 10 −5 M; b ). Passive pressure-diameter curves are also shown for easy comparison (black, trained; red, sedentary; a , b ). Mean values were used from 2-way ANOVA tests with Tukey paired comparisons: * p

    Techniques Used:

    Exercise training-induced remodeling of rat intramural coronary arterioles: a schematic representation. Wall sections of exercised and sedentary male rat coronary arteriole segments in spontaneous myogenic tone without ( a ) and with ( b , c ) inhibitors. a Coronary arterioles in control myogenic tone at 120 mm Hg intraluminal pressure and with the passive diameter. b The effect of the NO inhibitor N ω -nitro-L-arginine (in the presence of prostanoids) reveals an increased (NO-mediated) dilation effect in myogenic tone at 30 mm Hg intraluminal pressure in the coronary arterioles of exercised rats. c The effect of the cyclo-oxygenase inhibitor INDO (in the presence of NO) reveals the presence of endogenous constrictor prostanoids at 120 mm Hg intraluminal pressure in the coronary arterioles of sedentary animals, which is missing in the exercised group.
    Figure Legend Snippet: Exercise training-induced remodeling of rat intramural coronary arterioles: a schematic representation. Wall sections of exercised and sedentary male rat coronary arteriole segments in spontaneous myogenic tone without ( a ) and with ( b , c ) inhibitors. a Coronary arterioles in control myogenic tone at 120 mm Hg intraluminal pressure and with the passive diameter. b The effect of the NO inhibitor N ω -nitro-L-arginine (in the presence of prostanoids) reveals an increased (NO-mediated) dilation effect in myogenic tone at 30 mm Hg intraluminal pressure in the coronary arterioles of exercised rats. c The effect of the cyclo-oxygenase inhibitor INDO (in the presence of NO) reveals the presence of endogenous constrictor prostanoids at 120 mm Hg intraluminal pressure in the coronary arterioles of sedentary animals, which is missing in the exercised group.

    Techniques Used:

    4) Product Images from "Nitric Oxide and Hydrogen Peroxide Production are Involved in Systemic Drought Tolerance Induced by 2R,3R-Butanediol in Arabidopsis thaliana"

    Article Title: Nitric Oxide and Hydrogen Peroxide Production are Involved in Systemic Drought Tolerance Induced by 2R,3R-Butanediol in Arabidopsis thaliana

    Journal: The Plant Pathology Journal

    doi: 10.5423/PPJ.OA.07.2013.0069

    Survival of  Arabidopsis  Col-0 plants through induced drought resistance generated by 2R,3R-butanediol (2,3-B) in the presence and absence of chemicals to impair NO effects.  Arabidopsis  plants were grown in microtiter plates containing half-strength MS solid medium on sterile Whatman filter papers. Roots were treated with MES buffer as a control, or with 100 μM 2R,3R-butanediol in MES buffer in the presence or absence of 0.2 mM Nω-nitro- l -arginine (L-NNA), and 0.2 mM 2-phenyl-4,4,5,5-tetrmethyl-limidazolinone-1-oxyl-3-oxide (PTIO). The plants were exposed to drought stress after 3 days by transferring them to open Petri dishes, and wilted plants were counted after re-hydration. Three independent experiments were performed with at least 20 plants/treatment. Different letters indicate significant differences between treatments according to the least significant differences test at  P
    Figure Legend Snippet: Survival of Arabidopsis Col-0 plants through induced drought resistance generated by 2R,3R-butanediol (2,3-B) in the presence and absence of chemicals to impair NO effects. Arabidopsis plants were grown in microtiter plates containing half-strength MS solid medium on sterile Whatman filter papers. Roots were treated with MES buffer as a control, or with 100 μM 2R,3R-butanediol in MES buffer in the presence or absence of 0.2 mM Nω-nitro- l -arginine (L-NNA), and 0.2 mM 2-phenyl-4,4,5,5-tetrmethyl-limidazolinone-1-oxyl-3-oxide (PTIO). The plants were exposed to drought stress after 3 days by transferring them to open Petri dishes, and wilted plants were counted after re-hydration. Three independent experiments were performed with at least 20 plants/treatment. Different letters indicate significant differences between treatments according to the least significant differences test at P

    Techniques Used: Generated, Mass Spectrometry, Transferring

    Inhibition by tungstate (Tug) or NG-nitro- l -arginine methyl ester (L-NAME) of NO production stimulated by 2R,3R-butanediol (2,3-B). Epidermal cells in a leaf peel were incubated with DAF2-DA in MES buffer for 2 h prior to exposure to 100 μM 2R,3R-butanediol (2,3-B) in the absence or presence of L-NAME (200 μM) or tug (100 μM). Fluorescence was measured using a confocal microscope, and data are displayed as mean pixel intensities and associated standard deviations. A total of 50–100 guard cells were observed per treatment of three independent replicates. Different letters indicate significant difference between treatments according to the least significant differences test at  P
    Figure Legend Snippet: Inhibition by tungstate (Tug) or NG-nitro- l -arginine methyl ester (L-NAME) of NO production stimulated by 2R,3R-butanediol (2,3-B). Epidermal cells in a leaf peel were incubated with DAF2-DA in MES buffer for 2 h prior to exposure to 100 μM 2R,3R-butanediol (2,3-B) in the absence or presence of L-NAME (200 μM) or tug (100 μM). Fluorescence was measured using a confocal microscope, and data are displayed as mean pixel intensities and associated standard deviations. A total of 50–100 guard cells were observed per treatment of three independent replicates. Different letters indicate significant difference between treatments according to the least significant differences test at P

    Techniques Used: Inhibition, Incubation, Fluorescence, Microscopy

    5) Product Images from "Grape-Derived Polyphenols Improve Aging-Related Endothelial Dysfunction in Rat Mesenteric Artery: Role of Oxidative Stress and the Angiotensin System"

    Article Title: Grape-Derived Polyphenols Improve Aging-Related Endothelial Dysfunction in Rat Mesenteric Artery: Role of Oxidative Stress and the Angiotensin System

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0032039

    Acetylcholine-induced NO- and EDHF-mediated relaxations decrease with increasing age. Mesenteric artery rings from young (16-week old), mature-adult (25-week old) and middle-aged (46-week old) rats were contracted with phenylephrine (1 µM) in the presence of indomethacin (10 µM, to inhibit the formation of prostanoids) and (a) charybdotoxin (CTX, 100 nM) plus apamin (APA, 100 nM) to inhibit the participation of EDHF, or (b) N ω -nitro-L-arginine (L-NA, 300 µM) to rule out the formation of NO before a concentration-relaxation curve to acetylcholine was constructed. Results are shown as mean ± SEM of 5 to 6 different rats. * P
    Figure Legend Snippet: Acetylcholine-induced NO- and EDHF-mediated relaxations decrease with increasing age. Mesenteric artery rings from young (16-week old), mature-adult (25-week old) and middle-aged (46-week old) rats were contracted with phenylephrine (1 µM) in the presence of indomethacin (10 µM, to inhibit the formation of prostanoids) and (a) charybdotoxin (CTX, 100 nM) plus apamin (APA, 100 nM) to inhibit the participation of EDHF, or (b) N ω -nitro-L-arginine (L-NA, 300 µM) to rule out the formation of NO before a concentration-relaxation curve to acetylcholine was constructed. Results are shown as mean ± SEM of 5 to 6 different rats. * P

    Techniques Used: Concentration Assay, Construct

    The RWPs-induced Improvement of endothelial dysfunction in the mesenteric artery persists after 7 and 14-day washout periods. Middle-aged rats (46-week old) received RWPs (100 mg/kg/day) in the drinking water for either 21 days or 14 days followed by a 7-day washout period (a, b), and for 28 days or 14 days followed by a 14-day washout period (c, d). NO-mediated relaxations were determined in rings contracted with phenylephrine (1 µM) in the presence of indomethacin (10 µM) and charybdotoxin (CTX, 100 nM) plus apamin (APA, 100 nM) to inhibit the participation of prostanoids and EDHF, respectively (a, c). EDHF-mediated relaxations were recorded in the presence of indomethacin (10 µM) and N ω -nitro-L-arginine (L-NA, 300 µM) to rule out the participation of prostanoids and NO, respectively (b, d). Results are shown as means ± SEM of 5 to 6 rats. * P
    Figure Legend Snippet: The RWPs-induced Improvement of endothelial dysfunction in the mesenteric artery persists after 7 and 14-day washout periods. Middle-aged rats (46-week old) received RWPs (100 mg/kg/day) in the drinking water for either 21 days or 14 days followed by a 7-day washout period (a, b), and for 28 days or 14 days followed by a 14-day washout period (c, d). NO-mediated relaxations were determined in rings contracted with phenylephrine (1 µM) in the presence of indomethacin (10 µM) and charybdotoxin (CTX, 100 nM) plus apamin (APA, 100 nM) to inhibit the participation of prostanoids and EDHF, respectively (a, c). EDHF-mediated relaxations were recorded in the presence of indomethacin (10 µM) and N ω -nitro-L-arginine (L-NA, 300 µM) to rule out the participation of prostanoids and NO, respectively (b, d). Results are shown as means ± SEM of 5 to 6 rats. * P

    Techniques Used:

    Intake of RWPs improves blunted NO- and EDHF-mediated relaxations in the mesenteric artery of middle-aged rats. Young (16-week old) and middle-aged (46-week old) control rats received solvent (ethanol, 3% v/v) in the drinking water and middle-aged treated rats received RWPs (100 mg/kg/day) for either 7 (a, b) or 14 days before sacrifice (c, d). NO-mediated relaxations were determined in rings contracted with phenylephrine (1 µM) in the presence of indomethacin (10 µM) and charybdotoxin (CTX, 100 nM) plus apamin (APA, 100 nM) to inhibit the participation of prostanoids and EDHF, respectively (a, c). EDHF-mediated relaxations were determined in the presence of indomethacin (10 µM) and N ω -nitro-L-arginine (L-NA, 300 µM) to rule out the participation of prostanoids and NO, respectively (b, d). Results are shown as mean ± SEM of 5 to 6 different rats. * P
    Figure Legend Snippet: Intake of RWPs improves blunted NO- and EDHF-mediated relaxations in the mesenteric artery of middle-aged rats. Young (16-week old) and middle-aged (46-week old) control rats received solvent (ethanol, 3% v/v) in the drinking water and middle-aged treated rats received RWPs (100 mg/kg/day) for either 7 (a, b) or 14 days before sacrifice (c, d). NO-mediated relaxations were determined in rings contracted with phenylephrine (1 µM) in the presence of indomethacin (10 µM) and charybdotoxin (CTX, 100 nM) plus apamin (APA, 100 nM) to inhibit the participation of prostanoids and EDHF, respectively (a, c). EDHF-mediated relaxations were determined in the presence of indomethacin (10 µM) and N ω -nitro-L-arginine (L-NA, 300 µM) to rule out the participation of prostanoids and NO, respectively (b, d). Results are shown as mean ± SEM of 5 to 6 different rats. * P

    Techniques Used:

    Related Articles

    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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