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  • 99
    Thermo Fisher nacl
    CD spectra of the artificial WSTF PHD_EL5 RING finger and its five mutants. Spectra of 25 μM samples were collected in 20 mM <t>Tris-HCl</t> (pH 6.9), 50 mM <t>NaCl,</t> 1 mM dithiothreitol, and 50 μM ZnCl 2 at room temperature. (1) K4R, (2) K8R, (3) K9R, (4) K14R, and (5) K23R are denoted by solid lines, and the dotted line displays the wild-type.
    Nacl, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 14326 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Thermo Fisher 5m nacl
    CD spectra of the artificial WSTF PHD_EL5 RING finger and its five mutants. Spectra of 25 μM samples were collected in 20 mM <t>Tris-HCl</t> (pH 6.9), 50 mM <t>NaCl,</t> 1 mM dithiothreitol, and 50 μM ZnCl 2 at room temperature. (1) K4R, (2) K8R, (3) K9R, (4) K14R, and (5) K23R are denoted by solid lines, and the dotted line displays the wild-type.
    5m Nacl, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 53 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    nacl  (3M Co)
    92
    3M Co nacl
    CD spectra of the artificial WSTF PHD_EL5 RING finger and its five mutants. Spectra of 25 μM samples were collected in 20 mM <t>Tris-HCl</t> (pH 6.9), 50 mM <t>NaCl,</t> 1 mM dithiothreitol, and 50 μM ZnCl 2 at room temperature. (1) K4R, (2) K8R, (3) K9R, (4) K14R, and (5) K23R are denoted by solid lines, and the dotted line displays the wild-type.
    Nacl, supplied by 3M Co, used in various techniques. Bioz Stars score: 92/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Amresco nacl
    Figure 1. Expression patterns with functional convergence of OsAP2 and OsWRKY24 in rice. (A) In the RiceFREND database, OsAP2 (Os10g41330) marked as 1 is presented to be co-expressed with OsWRKY24 (Os01g61080) marked as 2. The red and blue circles represent plant-pathogen interaction and the phosphatidylinositol signaling system, respectively (in KEGG pathway). (B) Expression of OsAP2 and OsWRKY24 under different abiotic stress conditions. Rice seedlings were germinated and incubated on 1/2 MS media for 10 days and transferred to test tubes containing water (mock), <t>ABA</t> (60 µM; Sigma-A4906), PEG6000 (15%; Alfa Aesar-A17541) and <t>NaCl</t> (150 mM; Amresco-0241) solution, respectively. The plants were grown for 2 days under 12 h-light photoperiod at a constant temperature of 28°C, and each sample was harvested at 3 h, 12 h and 48 h time points. Actin ( Act ) with exogenous brassinolide treatment (10 −8 and 10 −6  M; Sigma-E1641). Leaves of transgenic rice plants expressing OsAP2 and OsWRKY24 under the control of OsBUL1 ) were used together with those of WT control, TNG67. Values represent mean±SD ( n ≥15). Data that is significantly different from the corresponding control is indicated (* p
    Nacl, supplied by Amresco, used in various techniques. Bioz Stars score: 92/100, based on 387 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Applichem nacl
    Figure 1. Expression patterns with functional convergence of OsAP2 and OsWRKY24 in rice. (A) In the RiceFREND database, OsAP2 (Os10g41330) marked as 1 is presented to be co-expressed with OsWRKY24 (Os01g61080) marked as 2. The red and blue circles represent plant-pathogen interaction and the phosphatidylinositol signaling system, respectively (in KEGG pathway). (B) Expression of OsAP2 and OsWRKY24 under different abiotic stress conditions. Rice seedlings were germinated and incubated on 1/2 MS media for 10 days and transferred to test tubes containing water (mock), <t>ABA</t> (60 µM; Sigma-A4906), PEG6000 (15%; Alfa Aesar-A17541) and <t>NaCl</t> (150 mM; Amresco-0241) solution, respectively. The plants were grown for 2 days under 12 h-light photoperiod at a constant temperature of 28°C, and each sample was harvested at 3 h, 12 h and 48 h time points. Actin ( Act ) with exogenous brassinolide treatment (10 −8 and 10 −6  M; Sigma-E1641). Leaves of transgenic rice plants expressing OsAP2 and OsWRKY24 under the control of OsBUL1 ) were used together with those of WT control, TNG67. Values represent mean±SD ( n ≥15). Data that is significantly different from the corresponding control is indicated (* p
    Nacl, supplied by Applichem, used in various techniques. Bioz Stars score: 92/100, based on 340 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Applygen Technologies nacl
    Figure 1. Expression patterns with functional convergence of OsAP2 and OsWRKY24 in rice. (A) In the RiceFREND database, OsAP2 (Os10g41330) marked as 1 is presented to be co-expressed with OsWRKY24 (Os01g61080) marked as 2. The red and blue circles represent plant-pathogen interaction and the phosphatidylinositol signaling system, respectively (in KEGG pathway). (B) Expression of OsAP2 and OsWRKY24 under different abiotic stress conditions. Rice seedlings were germinated and incubated on 1/2 MS media for 10 days and transferred to test tubes containing water (mock), <t>ABA</t> (60 µM; Sigma-A4906), PEG6000 (15%; Alfa Aesar-A17541) and <t>NaCl</t> (150 mM; Amresco-0241) solution, respectively. The plants were grown for 2 days under 12 h-light photoperiod at a constant temperature of 28°C, and each sample was harvested at 3 h, 12 h and 48 h time points. Actin ( Act ) with exogenous brassinolide treatment (10 −8 and 10 −6  M; Sigma-E1641). Leaves of transgenic rice plants expressing OsAP2 and OsWRKY24 under the control of OsBUL1 ) were used together with those of WT control, TNG67. Values represent mean±SD ( n ≥15). Data that is significantly different from the corresponding control is indicated (* p
    Nacl, supplied by Applygen Technologies, used in various techniques. Bioz Stars score: 92/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Avantor nacl
    Total body cortisol—comparison of HP proportion of total body cortisol and MGGF proportion of total body cortisol; HP = hepatopancreas; MGGF = sum of mantle, gills, gonads, foot; total cortisol = hepatopancreas, mantle, gills, gonads, foot; CuCl 2 = copper (II) chloride treatment; <t>NaCl</t> = sodium chloride treatment; A: pie charts are showing the distribution of total body cortisol between HP and MGGF. B: Bars are showing mean ± standard deviation of total body cortisol per mussel; *** are showing highly significant increase in the difference between HP and MGGF; CNF, n = 10; CF, n = 10; copper (II) chloride, n = 20; sodium chloride, n = 20
    Nacl, supplied by Avantor, used in various techniques. Bioz Stars score: 92/100, based on 496 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    B. Braun nacl
    Total body cortisol—comparison of HP proportion of total body cortisol and MGGF proportion of total body cortisol; HP = hepatopancreas; MGGF = sum of mantle, gills, gonads, foot; total cortisol = hepatopancreas, mantle, gills, gonads, foot; CuCl 2 = copper (II) chloride treatment; <t>NaCl</t> = sodium chloride treatment; A: pie charts are showing the distribution of total body cortisol between HP and MGGF. B: Bars are showing mean ± standard deviation of total body cortisol per mussel; *** are showing highly significant increase in the difference between HP and MGGF; CNF, n = 10; CF, n = 10; copper (II) chloride, n = 20; sodium chloride, n = 20
    Nacl, supplied by B. Braun, used in various techniques. Bioz Stars score: 92/100, based on 200 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Baxter Healthcare nacl
    Total body cortisol—comparison of HP proportion of total body cortisol and MGGF proportion of total body cortisol; HP = hepatopancreas; MGGF = sum of mantle, gills, gonads, foot; total cortisol = hepatopancreas, mantle, gills, gonads, foot; CuCl 2 = copper (II) chloride treatment; <t>NaCl</t> = sodium chloride treatment; A: pie charts are showing the distribution of total body cortisol between HP and MGGF. B: Bars are showing mean ± standard deviation of total body cortisol per mussel; *** are showing highly significant increase in the difference between HP and MGGF; CNF, n = 10; CF, n = 10; copper (II) chloride, n = 20; sodium chloride, n = 20
    Nacl, supplied by Baxter Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Becton Dickinson nacl
    Total body cortisol—comparison of HP proportion of total body cortisol and MGGF proportion of total body cortisol; HP = hepatopancreas; MGGF = sum of mantle, gills, gonads, foot; total cortisol = hepatopancreas, mantle, gills, gonads, foot; CuCl 2 = copper (II) chloride treatment; <t>NaCl</t> = sodium chloride treatment; A: pie charts are showing the distribution of total body cortisol between HP and MGGF. B: Bars are showing mean ± standard deviation of total body cortisol per mussel; *** are showing highly significant increase in the difference between HP and MGGF; CNF, n = 10; CF, n = 10; copper (II) chloride, n = 20; sodium chloride, n = 20
    Nacl, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 1101 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bio Basic Canada nacl
    Total body cortisol—comparison of HP proportion of total body cortisol and MGGF proportion of total body cortisol; HP = hepatopancreas; MGGF = sum of mantle, gills, gonads, foot; total cortisol = hepatopancreas, mantle, gills, gonads, foot; CuCl 2 = copper (II) chloride treatment; <t>NaCl</t> = sodium chloride treatment; A: pie charts are showing the distribution of total body cortisol between HP and MGGF. B: Bars are showing mean ± standard deviation of total body cortisol per mussel; *** are showing highly significant increase in the difference between HP and MGGF; CNF, n = 10; CF, n = 10; copper (II) chloride, n = 20; sodium chloride, n = 20
    Nacl, supplied by Bio Basic Canada, used in various techniques. Bioz Stars score: 92/100, based on 72 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    BioShop nacl
    Total body cortisol—comparison of HP proportion of total body cortisol and MGGF proportion of total body cortisol; HP = hepatopancreas; MGGF = sum of mantle, gills, gonads, foot; total cortisol = hepatopancreas, mantle, gills, gonads, foot; CuCl 2 = copper (II) chloride treatment; <t>NaCl</t> = sodium chloride treatment; A: pie charts are showing the distribution of total body cortisol between HP and MGGF. B: Bars are showing mean ± standard deviation of total body cortisol per mussel; *** are showing highly significant increase in the difference between HP and MGGF; CNF, n = 10; CF, n = 10; copper (II) chloride, n = 20; sodium chloride, n = 20
    Nacl, supplied by BioShop, used in various techniques. Bioz Stars score: 92/100, based on 111 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Carl Roth GmbH nacl
    UBC responses to <t>NaCl,</t> <t>ATP</t> and denatonium. Sequential stimulation with NaCl (50 mM on top of baseline concentrations in Tyrode), ATP (0.5 mM) and denatonium (25 mM); sequence of stimuli was changed between experiments ( N = 37). Graphs show representative recordings of changes in Calcium Orange® fluorescence in single cholinergic (eGFP + ) UBC. Experiments were performed during continuous superfusion with Tyrode III buffer Stimuli were added under continuous flow of Tyrode III into the chamber, so that indicated concentrations were reached initially and then washed out. Y-Axis depicts arbitrary units (AU) of Calcium Orange® fluorescence.
    Nacl, supplied by Carl Roth GmbH, used in various techniques. Bioz Stars score: 92/100, based on 530 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    nacl  (Difco)
    92
    Difco nacl
    Morphologies of Vibrio vulnificus ATCC <t>33815</t> incubated in artificial sea water (pH 6) microcosms with varying concentrations of <t>NaCl</t> at 4 °C on the seventh day. ( A , B ) Bacterial cells in a microcosm containing 0.75% NaCl; ( C , D ) cells in a microcosm amended with 5% NaCl; ( E , F ) cells in a microcosm amended with 10% NaCl; and ( G , H ) cells in a microcosm amended with 30% NaCl. Arrows point out the collapse of cytoplasmic spaces, which resulted in the development of gaps between the cell membrane and the peripheral membrane
    Nacl, supplied by Difco, used in various techniques. Bioz Stars score: 92/100, based on 346 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    EM Science Inc nacl
    Competitive interaction between TPrA and DEA. ( A ) Dependence of TPrA block on [DEA]. Current traces recorded from a single BTX-activated rat skeletal muscle sodium channel under control conditions (symmetrical 200 mM <t>NaCl,</t> 10 mM <t>MOPS,</t> pH 7.0). The upper
    Nacl, supplied by EM Science Inc, used in various techniques. Bioz Stars score: 92/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Euromedex nacl
    Competitive interaction between TPrA and DEA. ( A ) Dependence of TPrA block on [DEA]. Current traces recorded from a single BTX-activated rat skeletal muscle sodium channel under control conditions (symmetrical 200 mM <t>NaCl,</t> 10 mM <t>MOPS,</t> pH 7.0). The upper
    Nacl, supplied by Euromedex, used in various techniques. Bioz Stars score: 92/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Fisher Scientific nacl
    Adsorption tests performed over 6 h in <t>TSB</t> media containing <t>NaCl</t> (A), adjusted to different pH values (B) and containing the detergents Lutensol AO 7 (C) or SDS (D). Graphs show the number of free (unattached extracellular) phages. All experiments were carried out twice and in duplicates and mean values and standard deviations are shown.
    Nacl, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 93/100, based on 906 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Fresenius Kabi nacl
    Infusion profiles. Infusion lines were perfused with a biopharmaceutical (‘infusate’) for 95 min at 2 mL/min followed by flushing of the line with 0.9% <t>NaCl</t> at the same rate until the protein concentration dropped below the lower limit of quantitation (LLOQ) (0.04 g/L) ( a ). Alternatively, the infusion line was filled with a biopharmaceutical and flushed immediately ( b , c ). Mean ± SD cumulative wash-out ( b ) for an experimental IgG1 (3.75 mg/mL). Concentration profiles (mean ± SD) with or without (w/o) introducing an air bubble between the infusate and the flushing solution ( b ). Mean ± SD cumulative wash-out for different <t>biopharmaceuticals</t> at different concentrations and at different perfusion rates ( c ). The dots in c represent the theoretical cumulative wash-out if it is assumed that the samples with a concentration
    Nacl, supplied by Fresenius Kabi, used in various techniques. Bioz Stars score: 93/100, based on 83 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    FUJIFILM nacl
    A. Reporter assay of virF promoter activity in an hns mutant . An hns deletion mutant of S. sonnei strain MS4841 carrying virFTL-lacZ (striped bars) was grown in <t>YENB</t> media with or without 150 mM <t>NaCl</t> were subjected to the β-galactosidase assay. For a comparison of activities, the data from Figure 1C, Graph 1, which was derived from simultaneous assays, is indicated by three solid bars on the left side of the graph. Strain and concentration of NaCl are indicated at the bottom of the graph as follows: Wt, wild-type strain (solid bars); hns , hns deletion mutant (striped bars); YENB medium, 0 (white bars); YENB medium with 150 mM NaCl, 150 mM (gray bars). B. Western blot analysis of H-NS and CpxR expression . An overnight LB culture of MS390 at 30°C was inoculated into fresh media and then the cells were cultured until they reached mid-log phase ( A 600 = 0.8). Media, temperature (YENB at 37°C; LB at 30°C and 37°C) and the concentration of NaCl are indicated on the top of the panel. Antibodies used for detection are indicated on the right side of the panels. A cross-reactive unknown protein detected by the anti-H-NS antiserum was used as a loding control.
    Nacl, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 92/100, based on 777 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    GE Healthcare nacl
    Cofactor loss promotes dissociation of T1. ( A ) Gel filtration assays of recombinant T1 and T2 proteins in buffer containing 150 mM <t>NaCl,</t> 100 mM <t>Tris-HCl</t> pH 7.5, 1 mM DTT, supplemented with 5 μM ZnSO 4 (top panels) or 5 mM EDTA (bottom panels). ‘D’ and ‘M’ indicate the elution volume corresponding to the size of a dimer (150 kDa) or a monomer (75 kDa), respectively ( B ) Gel filtration assays in buffer supplemented with 5 μM ZnSO 4 of WT, double mutant T1-SY (Cys308Ser and His359Tyr) and triple mutant T1-SYY (containing the additional mutation His488Tyr). Pictures on the left are based on the E. coli ThrRS structure (PDB code 1QF6) and represent the zinc-binding pocket showing in green the zinc coordination residues and in red residues mutated in the SY and SYY proteins. ( C ) Diagram showing the position of insertions b and c of T1 and insertion a of T2.
    Nacl, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 94/100, based on 17206 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    HiMedia Laboratories nacl
    Evaluation of contribution of YEM medium components for silver nanoparticles generation upon autoclaving with AgNO 3 . a : Alteration in color of 0.25 mM AgNO 3 upon autoclaving with different components of YEM medium namely, yeast extract (YE), mannitol, <t>MgSO4,</t> <t>NaCl</t> and K2HPO4 medium individually. b : UV-Vis absorption spectra of 0.25 mM AgNO 3 after autoclaving with YEM medium, yeast extract, mannitol, MgSO4, NaCl and K2HPO4.
    Nacl, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 96 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Hospira nacl
    Evaluation of contribution of YEM medium components for silver nanoparticles generation upon autoclaving with AgNO 3 . a : Alteration in color of 0.25 mM AgNO 3 upon autoclaving with different components of YEM medium namely, yeast extract (YE), mannitol, <t>MgSO4,</t> <t>NaCl</t> and K2HPO4 medium individually. b : UV-Vis absorption spectra of 0.25 mM AgNO 3 after autoclaving with YEM medium, yeast extract, mannitol, MgSO4, NaCl and K2HPO4.
    Nacl, supplied by Hospira, used in various techniques. Bioz Stars score: 92/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    CD spectra of the artificial WSTF PHD_EL5 RING finger and its five mutants. Spectra of 25 μM samples were collected in 20 mM Tris-HCl (pH 6.9), 50 mM NaCl, 1 mM dithiothreitol, and 50 μM ZnCl 2 at room temperature. (1) K4R, (2) K8R, (3) K9R, (4) K14R, and (5) K23R are denoted by solid lines, and the dotted line displays the wild-type.

    Journal: Scientific Reports

    Article Title: Structural model of ubiquitin transfer onto an artificial RING finger as an E3 ligase

    doi: 10.1038/srep06574

    Figure Lengend Snippet: CD spectra of the artificial WSTF PHD_EL5 RING finger and its five mutants. Spectra of 25 μM samples were collected in 20 mM Tris-HCl (pH 6.9), 50 mM NaCl, 1 mM dithiothreitol, and 50 μM ZnCl 2 at room temperature. (1) K4R, (2) K8R, (3) K9R, (4) K14R, and (5) K23R are denoted by solid lines, and the dotted line displays the wild-type.

    Article Snippet: The peptides dissolved in 1 ml of 8 M guanidine-HCl were dialyzed against degassed Solution A (20 mM Tris-HCl (pH 6.9), 50 mM NaCl, 1 mM dithiothreitol, 50 μM ZnCl2 ) overnight at 4°C using a Slide-A-Lyzer dialysis cassette (Thermo scientific, Rockford, IL, USA) as described previously .

    Techniques:

    CaM directly interacts with DR5 in DR5 death domain in a Ca 2+ -dependent manner. A , CaM pull-down of purified DR5 cytoplasmic region (DR5 CR) and DR5 death domain (DR5 DD) in a 50 m m Tris, pH 7.6, 120 m m NaCl, 1% Brij buffer with 1 m m Ca 2+ , or 2 m m EGTA.

    Journal: The Journal of Biological Chemistry

    Article Title: Characterization of the Interactions between Calmodulin and Death Receptor 5 in Triple-negative and Estrogen Receptor-positive Breast Cancer Cells

    doi: 10.1074/jbc.M116.727727

    Figure Lengend Snippet: CaM directly interacts with DR5 in DR5 death domain in a Ca 2+ -dependent manner. A , CaM pull-down of purified DR5 cytoplasmic region (DR5 CR) and DR5 death domain (DR5 DD) in a 50 m m Tris, pH 7.6, 120 m m NaCl, 1% Brij buffer with 1 m m Ca 2+ , or 2 m m EGTA.

    Article Snippet: ER-positive or triple-negative breast cancer cells were lysed using a 50 m m Tris-HCL pH 7.5, 150 m m NaCl, 1% Triton X-100, and 1:100 Halt protease inhibitor (Thermo Fisher Scientific, Waltham, MA) buffer at 4 °C for 30 min.

    Techniques: Chick Chorioallantoic Membrane Assay, Purification

    P 12 /T 26 as a substrate for nucleotide incorporation catalyzed by NS5pol. A , the primer/template double-stranded RNA, P 12 /T 26 is shown. B , shown is a single nucleotide primer extension assay without enzyme-RNA preincubation. The reaction was initiated at 37 °C by the addition of NS5pol (0.6 μ m ) to a mixture containing P 12 /T 26 (0.5 μ m ) and GTP (0.5 m m ) in the reaction buffer (40 m m Tris-Cl, pH 7.4, 23.8 m m NaCl, 3 m m DTT, 15% glycerol, and 2 m m MgCl 2 ); aliquots of the reaction were quenched by a formamide-EDTA solution at the indicated times. The samples were resolved on a 16% denaturing polyacrylamide gel. C , shown is the time course of 13-mer product formation from the experiment described in B . The data were fit to a single exponential, yielding a rate of 0.00110 ± 0.00003 s −1 and an amplitude of 0.460 ± 0.005 μ m .

    Journal: The Journal of Biological Chemistry

    Article Title: Characterization of the Elongation Complex of Dengue Virus RNA Polymerase

    doi: 10.1074/jbc.M110.162685

    Figure Lengend Snippet: P 12 /T 26 as a substrate for nucleotide incorporation catalyzed by NS5pol. A , the primer/template double-stranded RNA, P 12 /T 26 is shown. B , shown is a single nucleotide primer extension assay without enzyme-RNA preincubation. The reaction was initiated at 37 °C by the addition of NS5pol (0.6 μ m ) to a mixture containing P 12 /T 26 (0.5 μ m ) and GTP (0.5 m m ) in the reaction buffer (40 m m Tris-Cl, pH 7.4, 23.8 m m NaCl, 3 m m DTT, 15% glycerol, and 2 m m MgCl 2 ); aliquots of the reaction were quenched by a formamide-EDTA solution at the indicated times. The samples were resolved on a 16% denaturing polyacrylamide gel. C , shown is the time course of 13-mer product formation from the experiment described in B . The data were fit to a single exponential, yielding a rate of 0.00110 ± 0.00003 s −1 and an amplitude of 0.460 ± 0.005 μ m .

    Article Snippet: MgCl2 , EDTA, NaCl solutions, and Tris-Cl buffers were purchased from Ambion (Austin, TX).

    Techniques: Primer Extension Assay

    Figure 1. Expression patterns with functional convergence of OsAP2 and OsWRKY24 in rice. (A) In the RiceFREND database, OsAP2 (Os10g41330) marked as 1 is presented to be co-expressed with OsWRKY24 (Os01g61080) marked as 2. The red and blue circles represent plant-pathogen interaction and the phosphatidylinositol signaling system, respectively (in KEGG pathway). (B) Expression of OsAP2 and OsWRKY24 under different abiotic stress conditions. Rice seedlings were germinated and incubated on 1/2 MS media for 10 days and transferred to test tubes containing water (mock), ABA (60 µM; Sigma-A4906), PEG6000 (15%; Alfa Aesar-A17541) and NaCl (150 mM; Amresco-0241) solution, respectively. The plants were grown for 2 days under 12 h-light photoperiod at a constant temperature of 28°C, and each sample was harvested at 3 h, 12 h and 48 h time points. Actin ( Act ) with exogenous brassinolide treatment (10 −8 and 10 −6  M; Sigma-E1641). Leaves of transgenic rice plants expressing OsAP2 and OsWRKY24 under the control of OsBUL1 ) were used together with those of WT control, TNG67. Values represent mean±SD ( n ≥15). Data that is significantly different from the corresponding control is indicated (* p

    Journal: Plant Biotechnology

    Article Title: Overexpression of OsAP2 and OsWRKY24 in Arabidopsis results in reduction of plant size

    doi: 10.5511/plantbiotechnology.18.0508a

    Figure Lengend Snippet: Figure 1. Expression patterns with functional convergence of OsAP2 and OsWRKY24 in rice. (A) In the RiceFREND database, OsAP2 (Os10g41330) marked as 1 is presented to be co-expressed with OsWRKY24 (Os01g61080) marked as 2. The red and blue circles represent plant-pathogen interaction and the phosphatidylinositol signaling system, respectively (in KEGG pathway). (B) Expression of OsAP2 and OsWRKY24 under different abiotic stress conditions. Rice seedlings were germinated and incubated on 1/2 MS media for 10 days and transferred to test tubes containing water (mock), ABA (60 µM; Sigma-A4906), PEG6000 (15%; Alfa Aesar-A17541) and NaCl (150 mM; Amresco-0241) solution, respectively. The plants were grown for 2 days under 12 h-light photoperiod at a constant temperature of 28°C, and each sample was harvested at 3 h, 12 h and 48 h time points. Actin ( Act ) with exogenous brassinolide treatment (10 −8 and 10 −6  M; Sigma-E1641). Leaves of transgenic rice plants expressing OsAP2 and OsWRKY24 under the control of OsBUL1 ) were used together with those of WT control, TNG67. Values represent mean±SD ( n ≥15). Data that is significantly different from the corresponding control is indicated (* p

    Article Snippet: Since some genes of the AP2 family and WRKY family are known to play pivotal roles in the adaptation of plants to abiotic stresses including drought stress ( ; ), we also investigated expression patterns of the two genes under several abiotic stress-inducing conditions using abscisic acid (60 µM ABA; Sigma-A4906), polyethylene glycol (15% PEG6000; Alfa Aesar-A17541) and NaCl (150 mM; Amresco-0241).

    Techniques: Functional Assay, Expressing, Incubation, Mass Spectrometry, Activated Clotting Time Assay, Transgenic Assay

    Total body cortisol—comparison of HP proportion of total body cortisol and MGGF proportion of total body cortisol; HP = hepatopancreas; MGGF = sum of mantle, gills, gonads, foot; total cortisol = hepatopancreas, mantle, gills, gonads, foot; CuCl 2 = copper (II) chloride treatment; NaCl = sodium chloride treatment; A: pie charts are showing the distribution of total body cortisol between HP and MGGF. B: Bars are showing mean ± standard deviation of total body cortisol per mussel; *** are showing highly significant increase in the difference between HP and MGGF; CNF, n = 10; CF, n = 10; copper (II) chloride, n = 20; sodium chloride, n = 20

    Journal: Conservation Physiology

    Article Title: Does environmental stress affect cortisol biodistribution in freshwater mussels?

    doi: 10.1093/conphys/coz101

    Figure Lengend Snippet: Total body cortisol—comparison of HP proportion of total body cortisol and MGGF proportion of total body cortisol; HP = hepatopancreas; MGGF = sum of mantle, gills, gonads, foot; total cortisol = hepatopancreas, mantle, gills, gonads, foot; CuCl 2 = copper (II) chloride treatment; NaCl = sodium chloride treatment; A: pie charts are showing the distribution of total body cortisol between HP and MGGF. B: Bars are showing mean ± standard deviation of total body cortisol per mussel; *** are showing highly significant increase in the difference between HP and MGGF; CNF, n = 10; CF, n = 10; copper (II) chloride, n = 20; sodium chloride, n = 20

    Article Snippet: For the treatments, either CuCl2 (Carl Roth GmbH, Germany) or NaCl (crystal, J.T.BAKER, Netherlands) was added.

    Techniques: Standard Deviation

    UBC responses to NaCl, ATP and denatonium. Sequential stimulation with NaCl (50 mM on top of baseline concentrations in Tyrode), ATP (0.5 mM) and denatonium (25 mM); sequence of stimuli was changed between experiments ( N = 37). Graphs show representative recordings of changes in Calcium Orange® fluorescence in single cholinergic (eGFP + ) UBC. Experiments were performed during continuous superfusion with Tyrode III buffer Stimuli were added under continuous flow of Tyrode III into the chamber, so that indicated concentrations were reached initially and then washed out. Y-Axis depicts arbitrary units (AU) of Calcium Orange® fluorescence.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: ENaC in Cholinergic Brush Cells

    doi: 10.3389/fcell.2018.00089

    Figure Lengend Snippet: UBC responses to NaCl, ATP and denatonium. Sequential stimulation with NaCl (50 mM on top of baseline concentrations in Tyrode), ATP (0.5 mM) and denatonium (25 mM); sequence of stimuli was changed between experiments ( N = 37). Graphs show representative recordings of changes in Calcium Orange® fluorescence in single cholinergic (eGFP + ) UBC. Experiments were performed during continuous superfusion with Tyrode III buffer Stimuli were added under continuous flow of Tyrode III into the chamber, so that indicated concentrations were reached initially and then washed out. Y-Axis depicts arbitrary units (AU) of Calcium Orange® fluorescence.

    Article Snippet: Test stimuli and concentrations were denatonium benzoate (25 mM; Molekula, Munich, Germany), ATP (0.5 mM; Sigma-Aldrich/Merck, Darmstadt, Germany) and NaCl (1–150 mM; Carl Roth, Karlsruhe, Germany), and inhibitors and controls included the osmolarity control mannitol (1–150 mM; Sigma-Aldrich/Merck, Darmstadt, Germany) and the ENaC inhibitor amiloride (0.01–100 μM; Sigma-Aldrich/Merck, Darmstadt, Germany).

    Techniques: Sequencing, Fluorescence, Flow Cytometry

    Morphologies of Vibrio vulnificus ATCC 33815 incubated in artificial sea water (pH 6) microcosms with varying concentrations of NaCl at 4 °C on the seventh day. ( A , B ) Bacterial cells in a microcosm containing 0.75% NaCl; ( C , D ) cells in a microcosm amended with 5% NaCl; ( E , F ) cells in a microcosm amended with 10% NaCl; and ( G , H ) cells in a microcosm amended with 30% NaCl. Arrows point out the collapse of cytoplasmic spaces, which resulted in the development of gaps between the cell membrane and the peripheral membrane

    Journal: Food Science and Biotechnology

    Article Title: Effects of varying concentrations of sodium chloride and acidic conditions on the behavior of Vibrio parahaemolyticus and Vibrio vulnificus cold-starved in artificial sea water microcosms

    doi: 10.1007/s10068-017-0105-3

    Figure Lengend Snippet: Morphologies of Vibrio vulnificus ATCC 33815 incubated in artificial sea water (pH 6) microcosms with varying concentrations of NaCl at 4 °C on the seventh day. ( A , B ) Bacterial cells in a microcosm containing 0.75% NaCl; ( C , D ) cells in a microcosm amended with 5% NaCl; ( E , F ) cells in a microcosm amended with 10% NaCl; and ( G , H ) cells in a microcosm amended with 30% NaCl. Arrows point out the collapse of cytoplasmic spaces, which resulted in the development of gaps between the cell membrane and the peripheral membrane

    Article Snippet: V. parahaemolyticus ATCC 27969, V. parahaemolyticus ATCC 33844, and V. vulnificus ATCC 33815 in ASW were plating-counted on tryptic soy agar supplemented with 3% NaCl (TSA, Difco) and thiosulphate-citrate-bile salts-sucrose (TCBS, Difco).

    Techniques: Incubation

    Loss of culturability in Vibrio vulnificus ATCC 33815 incubated in artificial sea water microcosms (pH 4, 5, 6, and 7) containing ( A ) 0.75% NaCl or supplemented with ( B ) 5%, ( C ) 10%, and ( D ) 30% NaCl, respectively, at 4 °C for 30 days. These culutrabilities were enumerated on a nonselective medium (tryptic soy agar, TSA). The number of viable cells of V. vulnificus ATCC 33815 was measured via the fluorescent microscopic assay using the Live/Dead BacLight R bacterial viability kit. Filled circle , culturability at pH 7; open circle , viability at pH 7; filled triangle , culturability at pH 6; open triangle , viability at pH 6; filled square , culturability at pH 5; open square , viability at pH 5; filled diamond culturability at pH 4; open diamond , viability at pH 4

    Journal: Food Science and Biotechnology

    Article Title: Effects of varying concentrations of sodium chloride and acidic conditions on the behavior of Vibrio parahaemolyticus and Vibrio vulnificus cold-starved in artificial sea water microcosms

    doi: 10.1007/s10068-017-0105-3

    Figure Lengend Snippet: Loss of culturability in Vibrio vulnificus ATCC 33815 incubated in artificial sea water microcosms (pH 4, 5, 6, and 7) containing ( A ) 0.75% NaCl or supplemented with ( B ) 5%, ( C ) 10%, and ( D ) 30% NaCl, respectively, at 4 °C for 30 days. These culutrabilities were enumerated on a nonselective medium (tryptic soy agar, TSA). The number of viable cells of V. vulnificus ATCC 33815 was measured via the fluorescent microscopic assay using the Live/Dead BacLight R bacterial viability kit. Filled circle , culturability at pH 7; open circle , viability at pH 7; filled triangle , culturability at pH 6; open triangle , viability at pH 6; filled square , culturability at pH 5; open square , viability at pH 5; filled diamond culturability at pH 4; open diamond , viability at pH 4

    Article Snippet: V. parahaemolyticus ATCC 27969, V. parahaemolyticus ATCC 33844, and V. vulnificus ATCC 33815 in ASW were plating-counted on tryptic soy agar supplemented with 3% NaCl (TSA, Difco) and thiosulphate-citrate-bile salts-sucrose (TCBS, Difco).

    Techniques: Incubation

    Competitive interaction between TPrA and DEA. ( A ) Dependence of TPrA block on [DEA]. Current traces recorded from a single BTX-activated rat skeletal muscle sodium channel under control conditions (symmetrical 200 mM NaCl, 10 mM MOPS, pH 7.0). The upper

    Journal:

    Article Title: Trans-Channel Interactions in Batrachotoxin-Modified Rat Skeletal Muscle Sodium Channels: Kinetic Analysis of Mutual Inhibition between μ-Conotoxin GIIIA Derivatives and Amine Blockers

    doi: 10.1529/biophysj.108.138271

    Figure Lengend Snippet: Competitive interaction between TPrA and DEA. ( A ) Dependence of TPrA block on [DEA]. Current traces recorded from a single BTX-activated rat skeletal muscle sodium channel under control conditions (symmetrical 200 mM NaCl, 10 mM MOPS, pH 7.0). The upper

    Article Snippet: One to five microliters of incubated vesicles was pipetted into one well of a bilayer chamber containing a bathing solution of 200 mM NaCl (EM Science, Gibbstown, NJ), 10 mM MOPS (Sigma-Aldrich, Oakville, Ontario, Canada), 0.1 mM Na2 EDTA (Sigma-Aldrich), pH 7 (NaOH; BDH from VWR International, Edmonton, Alberta, Canada) in both wells.

    Techniques: Blocking Assay

    Concentration dependence of DEA block of single sodium channels, when either bound or unbound by R13E. Current traces recorded from a single BTX-activated rat skeletal muscle sodium channel under control conditions (symmetrical 200 mM NaCl, 10 mM MOPS,

    Journal:

    Article Title: Trans-Channel Interactions in Batrachotoxin-Modified Rat Skeletal Muscle Sodium Channels: Kinetic Analysis of Mutual Inhibition between μ-Conotoxin GIIIA Derivatives and Amine Blockers

    doi: 10.1529/biophysj.108.138271

    Figure Lengend Snippet: Concentration dependence of DEA block of single sodium channels, when either bound or unbound by R13E. Current traces recorded from a single BTX-activated rat skeletal muscle sodium channel under control conditions (symmetrical 200 mM NaCl, 10 mM MOPS,

    Article Snippet: One to five microliters of incubated vesicles was pipetted into one well of a bilayer chamber containing a bathing solution of 200 mM NaCl (EM Science, Gibbstown, NJ), 10 mM MOPS (Sigma-Aldrich, Oakville, Ontario, Canada), 0.1 mM Na2 EDTA (Sigma-Aldrich), pH 7 (NaOH; BDH from VWR International, Edmonton, Alberta, Canada) in both wells.

    Techniques: Concentration Assay, Blocking Assay

    Adsorption tests performed over 6 h in TSB media containing NaCl (A), adjusted to different pH values (B) and containing the detergents Lutensol AO 7 (C) or SDS (D). Graphs show the number of free (unattached extracellular) phages. All experiments were carried out twice and in duplicates and mean values and standard deviations are shown.

    Journal: Frontiers in Microbiology

    Article Title: Influence of Environmental Factors on Phage–Bacteria Interaction and on the Efficacy and Infectivity of Phage P100

    doi: 10.3389/fmicb.2016.01152

    Figure Lengend Snippet: Adsorption tests performed over 6 h in TSB media containing NaCl (A), adjusted to different pH values (B) and containing the detergents Lutensol AO 7 (C) or SDS (D). Graphs show the number of free (unattached extracellular) phages. All experiments were carried out twice and in duplicates and mean values and standard deviations are shown.

    Article Snippet: To test the short-time influence of chemicals on P100 stability in terms of infectivity, phages were incubated in TSB containing either 2 M NaCl (Fisher Scientific, Leics, UK), the detergents Lutensol AO 7 (BASF, Ludwigshafen, Germany) and sodium dodecyl sulfate (SDS; SIGMA-ALDRICH, Steinheim, Germany; 5% each) and TSB adjusted to the pH values of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 using 0.5 M HCl or NaOH (Merck, Darmstadt, Germany).

    Techniques: Adsorption

    Adsorption tests performed over 60 min indicating attachment (decreasing number of free phages) of phage P100 to L. monocytogenes in TSB containing NaCl (A), Lutensol AO 7 (B), TSB adjusted to pH values 3–11 (C), smear water and Fraser (D), and TSB containing SDS (E). All experiments were done twice and in duplicates and mean values and standard deviations were shown.

    Journal: Frontiers in Microbiology

    Article Title: Influence of Environmental Factors on Phage–Bacteria Interaction and on the Efficacy and Infectivity of Phage P100

    doi: 10.3389/fmicb.2016.01152

    Figure Lengend Snippet: Adsorption tests performed over 60 min indicating attachment (decreasing number of free phages) of phage P100 to L. monocytogenes in TSB containing NaCl (A), Lutensol AO 7 (B), TSB adjusted to pH values 3–11 (C), smear water and Fraser (D), and TSB containing SDS (E). All experiments were done twice and in duplicates and mean values and standard deviations were shown.

    Article Snippet: To test the short-time influence of chemicals on P100 stability in terms of infectivity, phages were incubated in TSB containing either 2 M NaCl (Fisher Scientific, Leics, UK), the detergents Lutensol AO 7 (BASF, Ludwigshafen, Germany) and sodium dodecyl sulfate (SDS; SIGMA-ALDRICH, Steinheim, Germany; 5% each) and TSB adjusted to the pH values of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 using 0.5 M HCl or NaOH (Merck, Darmstadt, Germany).

    Techniques: Adsorption

    Growth curves of uninfected (A) and infected (B,C; MOI = 10) L. monocytogenes in TSB and NaCl containing TSB media. Bacteria concentrations at the beginning of the infections were 10 7 CFU/ml (A,B) and 10 6 CFU/ml (C) . All NaCl concentrations (0–2 M) were tested. For the sake of clarity only the curves of growing bacteria were demonstrated while the others on base line level were not depicted. Moreover, one of four independent experiments is representatively shown.

    Journal: Frontiers in Microbiology

    Article Title: Influence of Environmental Factors on Phage–Bacteria Interaction and on the Efficacy and Infectivity of Phage P100

    doi: 10.3389/fmicb.2016.01152

    Figure Lengend Snippet: Growth curves of uninfected (A) and infected (B,C; MOI = 10) L. monocytogenes in TSB and NaCl containing TSB media. Bacteria concentrations at the beginning of the infections were 10 7 CFU/ml (A,B) and 10 6 CFU/ml (C) . All NaCl concentrations (0–2 M) were tested. For the sake of clarity only the curves of growing bacteria were demonstrated while the others on base line level were not depicted. Moreover, one of four independent experiments is representatively shown.

    Article Snippet: To test the short-time influence of chemicals on P100 stability in terms of infectivity, phages were incubated in TSB containing either 2 M NaCl (Fisher Scientific, Leics, UK), the detergents Lutensol AO 7 (BASF, Ludwigshafen, Germany) and sodium dodecyl sulfate (SDS; SIGMA-ALDRICH, Steinheim, Germany; 5% each) and TSB adjusted to the pH values of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 using 0.5 M HCl or NaOH (Merck, Darmstadt, Germany).

    Techniques: Infection

    Infusion profiles. Infusion lines were perfused with a biopharmaceutical (‘infusate’) for 95 min at 2 mL/min followed by flushing of the line with 0.9% NaCl at the same rate until the protein concentration dropped below the lower limit of quantitation (LLOQ) (0.04 g/L) ( a ). Alternatively, the infusion line was filled with a biopharmaceutical and flushed immediately ( b , c ). Mean ± SD cumulative wash-out ( b ) for an experimental IgG1 (3.75 mg/mL). Concentration profiles (mean ± SD) with or without (w/o) introducing an air bubble between the infusate and the flushing solution ( b ). Mean ± SD cumulative wash-out for different biopharmaceuticals at different concentrations and at different perfusion rates ( c ). The dots in c represent the theoretical cumulative wash-out if it is assumed that the samples with a concentration

    Journal: European Journal of Drug Metabolism and Pharmacokinetics

    Article Title: Potential Influence of Endothelial Adsorption on the Delayed Time to Maximum Concentration of Biopharmaceuticals

    doi: 10.1007/s13318-017-0430-1

    Figure Lengend Snippet: Infusion profiles. Infusion lines were perfused with a biopharmaceutical (‘infusate’) for 95 min at 2 mL/min followed by flushing of the line with 0.9% NaCl at the same rate until the protein concentration dropped below the lower limit of quantitation (LLOQ) (0.04 g/L) ( a ). Alternatively, the infusion line was filled with a biopharmaceutical and flushed immediately ( b , c ). Mean ± SD cumulative wash-out ( b ) for an experimental IgG1 (3.75 mg/mL). Concentration profiles (mean ± SD) with or without (w/o) introducing an air bubble between the infusate and the flushing solution ( b ). Mean ± SD cumulative wash-out for different biopharmaceuticals at different concentrations and at different perfusion rates ( c ). The dots in c represent the theoretical cumulative wash-out if it is assumed that the samples with a concentration

    Article Snippet: The dilutions of the biopharmaceuticals in 0.9% NaCl (Fresenius Kabi, Schelle, Belgium) were prepared by the LUMC Pharmacy.

    Techniques: Protein Concentration, Quantitation Assay, Concentration Assay

    A. Reporter assay of virF promoter activity in an hns mutant . An hns deletion mutant of S. sonnei strain MS4841 carrying virFTL-lacZ (striped bars) was grown in YENB media with or without 150 mM NaCl were subjected to the β-galactosidase assay. For a comparison of activities, the data from Figure 1C, Graph 1, which was derived from simultaneous assays, is indicated by three solid bars on the left side of the graph. Strain and concentration of NaCl are indicated at the bottom of the graph as follows: Wt, wild-type strain (solid bars); hns , hns deletion mutant (striped bars); YENB medium, 0 (white bars); YENB medium with 150 mM NaCl, 150 mM (gray bars). B. Western blot analysis of H-NS and CpxR expression . An overnight LB culture of MS390 at 30°C was inoculated into fresh media and then the cells were cultured until they reached mid-log phase ( A 600 = 0.8). Media, temperature (YENB at 37°C; LB at 30°C and 37°C) and the concentration of NaCl are indicated on the top of the panel. Antibodies used for detection are indicated on the right side of the panels. A cross-reactive unknown protein detected by the anti-H-NS antiserum was used as a loding control.

    Journal: BMC Microbiology

    Article Title: Involvement of RNA-binding protein Hfq in the osmotic-response regulation of invE gene expression in Shigella sonnei

    doi: 10.1186/1471-2180-9-110

    Figure Lengend Snippet: A. Reporter assay of virF promoter activity in an hns mutant . An hns deletion mutant of S. sonnei strain MS4841 carrying virFTL-lacZ (striped bars) was grown in YENB media with or without 150 mM NaCl were subjected to the β-galactosidase assay. For a comparison of activities, the data from Figure 1C, Graph 1, which was derived from simultaneous assays, is indicated by three solid bars on the left side of the graph. Strain and concentration of NaCl are indicated at the bottom of the graph as follows: Wt, wild-type strain (solid bars); hns , hns deletion mutant (striped bars); YENB medium, 0 (white bars); YENB medium with 150 mM NaCl, 150 mM (gray bars). B. Western blot analysis of H-NS and CpxR expression . An overnight LB culture of MS390 at 30°C was inoculated into fresh media and then the cells were cultured until they reached mid-log phase ( A 600 = 0.8). Media, temperature (YENB at 37°C; LB at 30°C and 37°C) and the concentration of NaCl are indicated on the top of the panel. Antibodies used for detection are indicated on the right side of the panels. A cross-reactive unknown protein detected by the anti-H-NS antiserum was used as a loding control.

    Article Snippet: YENB medium containing 150 mM NaCl (Wako Chemical, Tokyo Japan) was used as the physiological osmotic medium.

    Techniques: Reporter Assay, Activity Assay, Mutagenesis, Derivative Assay, Concentration Assay, Western Blot, Expressing, Cell Culture

    A. InvE expression in Δp invE:: p araBAD strain MS5512 . Δp invE:: p araBAD strain MS5512 and wild-type strain MS390 were grown overnight in LB medium containing chloramphenicol and 50 μM arabinose, washed twice with fresh LB medium, and then inoculated into YENB media containing increasing concentrations of arabinose and cultured at 37°C with or without 150 mM NaCl, as indicated. Strains (Δ pinvE::paraBAD , MS5512; Wt, wild-type strain MS390), concentration of NaCl (0 mM or 150 mM) and concentration of arabinose (0, 0.2, 0.5, 1.0 mM) are indicated above the panels. Primers and antibodies used in the experiments are indicated on the right side of the panels. B. Stability of InvE protein . Δ invE strain MS1632 carrying the expression plasmid pBAD-invE was grown in YENB media containing ampicillin and 100 μM arabinose, with or without 150 mM NaCl, at 37°C. When cultures reached an A 600 of 0.8, rifampicin was added. Cells were harvested at 10 min intervals for a period of 40 min. Whole cell cultures (10 μl) were analysed by Western blot using anti-InvE and -IpaB antibodies.

    Journal: BMC Microbiology

    Article Title: Involvement of RNA-binding protein Hfq in the osmotic-response regulation of invE gene expression in Shigella sonnei

    doi: 10.1186/1471-2180-9-110

    Figure Lengend Snippet: A. InvE expression in Δp invE:: p araBAD strain MS5512 . Δp invE:: p araBAD strain MS5512 and wild-type strain MS390 were grown overnight in LB medium containing chloramphenicol and 50 μM arabinose, washed twice with fresh LB medium, and then inoculated into YENB media containing increasing concentrations of arabinose and cultured at 37°C with or without 150 mM NaCl, as indicated. Strains (Δ pinvE::paraBAD , MS5512; Wt, wild-type strain MS390), concentration of NaCl (0 mM or 150 mM) and concentration of arabinose (0, 0.2, 0.5, 1.0 mM) are indicated above the panels. Primers and antibodies used in the experiments are indicated on the right side of the panels. B. Stability of InvE protein . Δ invE strain MS1632 carrying the expression plasmid pBAD-invE was grown in YENB media containing ampicillin and 100 μM arabinose, with or without 150 mM NaCl, at 37°C. When cultures reached an A 600 of 0.8, rifampicin was added. Cells were harvested at 10 min intervals for a period of 40 min. Whole cell cultures (10 μl) were analysed by Western blot using anti-InvE and -IpaB antibodies.

    Article Snippet: YENB medium containing 150 mM NaCl (Wako Chemical, Tokyo Japan) was used as the physiological osmotic medium.

    Techniques: Expressing, Cell Culture, Concentration Assay, Plasmid Preparation, Western Blot

    A. InvE and IpaB expression in the hfq deletion mutant . Wild-type strain MS390 and the hfq mutant strain MS4831 were cultured in YENB media with or without NaCl, and then subjected to Western blot analysis. Strains and concentration of NaCl are indicated above the panels. Antibodies used in the experiment are indicated on the right side of the panels. B. Effect of ectopic Hfq expression on InvE and IpaB in the hfq mutant . hfq deletion mutants carrying an Hfq expression plasmid or a control plasmid were subjected to Western blot analysis. Strains were grown in YENB medium containing ampicillin and IPTG, or YENB medium containing ampicillin, IPTG and 150 mM NaCl at 37°C, and then harvested. Strains, concentration of NaCl and plasmids (minus, pTrc99A; plus, pTrc-hfq) are indicated above the panel. Lane 1, wild-type strain MS390 grown in YENB medium; Lane 2, Δ hfq (pTrc99A) grown in YENB plus 0.1 mM IPTG; Lane 3, Δ hfq (pTrc-hfq) grown in YENB plus 0.1 mM IPTG; Lane 4, Δ hfq (pTrc99A) grown in YENB with 150 mM NaCl plus 1 mM IPTG; Lane 5, Δ hfq (pTrc-hfq) grown in YENB with 150 mM NaCl plus 1 mM IPTG.

    Journal: BMC Microbiology

    Article Title: Involvement of RNA-binding protein Hfq in the osmotic-response regulation of invE gene expression in Shigella sonnei

    doi: 10.1186/1471-2180-9-110

    Figure Lengend Snippet: A. InvE and IpaB expression in the hfq deletion mutant . Wild-type strain MS390 and the hfq mutant strain MS4831 were cultured in YENB media with or without NaCl, and then subjected to Western blot analysis. Strains and concentration of NaCl are indicated above the panels. Antibodies used in the experiment are indicated on the right side of the panels. B. Effect of ectopic Hfq expression on InvE and IpaB in the hfq mutant . hfq deletion mutants carrying an Hfq expression plasmid or a control plasmid were subjected to Western blot analysis. Strains were grown in YENB medium containing ampicillin and IPTG, or YENB medium containing ampicillin, IPTG and 150 mM NaCl at 37°C, and then harvested. Strains, concentration of NaCl and plasmids (minus, pTrc99A; plus, pTrc-hfq) are indicated above the panel. Lane 1, wild-type strain MS390 grown in YENB medium; Lane 2, Δ hfq (pTrc99A) grown in YENB plus 0.1 mM IPTG; Lane 3, Δ hfq (pTrc-hfq) grown in YENB plus 0.1 mM IPTG; Lane 4, Δ hfq (pTrc99A) grown in YENB with 150 mM NaCl plus 1 mM IPTG; Lane 5, Δ hfq (pTrc-hfq) grown in YENB with 150 mM NaCl plus 1 mM IPTG.

    Article Snippet: YENB medium containing 150 mM NaCl (Wako Chemical, Tokyo Japan) was used as the physiological osmotic medium.

    Techniques: Expressing, Mutagenesis, Cell Culture, Western Blot, Concentration Assay, Plasmid Preparation

    A. Stability of invE mRNA in low osmotic growth conditions . Pre-cultures were inoculated into 35 ml of fresh YENB media and then grown at 37°C with shaking. When cultures reached an A 600 of 0.8, rifampicin was added, then cells were harvested at 2 min intervals. Total RNA (100 ng) was used for RT-PCR analysis, and 10 μl of the amplified product was subjected to agarose gel electrophoresis. NaCl concentration (150 mM, 0 mM), strains (Wild-type strain MS390; Δ hfq , MS4831) and time after rifampicin treatment (0, 2, 4, 6, 8, or 32 min) are indicated above the panels. Primers used in the experiments are indicated on the right side of the panels. B. Decay curves of invE mRNAs . Total RNA (100 ng) was subjected to real-time PCR analysis. The amount of RNA was normalized to an internal control (6S RNA) and expression was expressed relative to expression at time 0, which was set as 1.0. The X-axis indicates time after rifampicin treatment (0 to 8 min). Presence or absence of 150 mM NaCl (plus, minus) and strains (Wt, wild-type strain MS390; Δ hfq , MS4831) are indicated on the right side of the graph.

    Journal: BMC Microbiology

    Article Title: Involvement of RNA-binding protein Hfq in the osmotic-response regulation of invE gene expression in Shigella sonnei

    doi: 10.1186/1471-2180-9-110

    Figure Lengend Snippet: A. Stability of invE mRNA in low osmotic growth conditions . Pre-cultures were inoculated into 35 ml of fresh YENB media and then grown at 37°C with shaking. When cultures reached an A 600 of 0.8, rifampicin was added, then cells were harvested at 2 min intervals. Total RNA (100 ng) was used for RT-PCR analysis, and 10 μl of the amplified product was subjected to agarose gel electrophoresis. NaCl concentration (150 mM, 0 mM), strains (Wild-type strain MS390; Δ hfq , MS4831) and time after rifampicin treatment (0, 2, 4, 6, 8, or 32 min) are indicated above the panels. Primers used in the experiments are indicated on the right side of the panels. B. Decay curves of invE mRNAs . Total RNA (100 ng) was subjected to real-time PCR analysis. The amount of RNA was normalized to an internal control (6S RNA) and expression was expressed relative to expression at time 0, which was set as 1.0. The X-axis indicates time after rifampicin treatment (0 to 8 min). Presence or absence of 150 mM NaCl (plus, minus) and strains (Wt, wild-type strain MS390; Δ hfq , MS4831) are indicated on the right side of the graph.

    Article Snippet: YENB medium containing 150 mM NaCl (Wako Chemical, Tokyo Japan) was used as the physiological osmotic medium.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Concentration Assay, Real-time Polymerase Chain Reaction, Expressing

    A. InvE and IpaB expression in different osmotic conditions . An overnight culture of strain MS390 at 30°C was inoculated into fresh YENB medium with or without osmolytes and then incubated at 37°C until mid-log phase ( A 600 = 0.8). Medium, osmolyte, and concentration are indicated at the top of the panel. Antibodies used for detection are indicated on the right of the panels. A cross-reactive unknown protein detected by the anti-InvE antiserum was used as a loading control for InvE Western blot analysis throughout this study. B . Expression of > invE and virF mRNA and InvE and IpaB protein expression in S. Sonnei . Total RNA (100 ng) and 10 μl of the indicate culture were subjected to analysis of mRNA and protein levels, respectively. The 6S RNA ssrS gene was used as control for RT-PCR. Primers and antibodies are indicated on the right side of the panels. Concentration of NaCl in the medium is indicated at top of the panel. C. Expression of invE and virF > promoter-driven reporter genes . Wild-type S. sonnei strain MS390 carrying the indicated reporter plasmids were subjected to a β-galactosidase assay: Graph 1, virFTL-lacZ translational fusion plasmid pHW848; Graph 2, invETx-lacZ transcriptional fusion plasmid pJM4320; Graph 3, invETL-lacZ translational fusion plasmid pJM4321. Concentration of NaCl is indicated at the bottom of the graphs. Details of the control experiments, indicated by black bars (NC)are described in methods.

    Journal: BMC Microbiology

    Article Title: Involvement of RNA-binding protein Hfq in the osmotic-response regulation of invE gene expression in Shigella sonnei

    doi: 10.1186/1471-2180-9-110

    Figure Lengend Snippet: A. InvE and IpaB expression in different osmotic conditions . An overnight culture of strain MS390 at 30°C was inoculated into fresh YENB medium with or without osmolytes and then incubated at 37°C until mid-log phase ( A 600 = 0.8). Medium, osmolyte, and concentration are indicated at the top of the panel. Antibodies used for detection are indicated on the right of the panels. A cross-reactive unknown protein detected by the anti-InvE antiserum was used as a loading control for InvE Western blot analysis throughout this study. B . Expression of > invE and virF mRNA and InvE and IpaB protein expression in S. Sonnei . Total RNA (100 ng) and 10 μl of the indicate culture were subjected to analysis of mRNA and protein levels, respectively. The 6S RNA ssrS gene was used as control for RT-PCR. Primers and antibodies are indicated on the right side of the panels. Concentration of NaCl in the medium is indicated at top of the panel. C. Expression of invE and virF > promoter-driven reporter genes . Wild-type S. sonnei strain MS390 carrying the indicated reporter plasmids were subjected to a β-galactosidase assay: Graph 1, virFTL-lacZ translational fusion plasmid pHW848; Graph 2, invETx-lacZ transcriptional fusion plasmid pJM4320; Graph 3, invETL-lacZ translational fusion plasmid pJM4321. Concentration of NaCl is indicated at the bottom of the graphs. Details of the control experiments, indicated by black bars (NC)are described in methods.

    Article Snippet: YENB medium containing 150 mM NaCl (Wako Chemical, Tokyo Japan) was used as the physiological osmotic medium.

    Techniques: Expressing, Incubation, Concentration Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation

    Cofactor loss promotes dissociation of T1. ( A ) Gel filtration assays of recombinant T1 and T2 proteins in buffer containing 150 mM NaCl, 100 mM Tris-HCl pH 7.5, 1 mM DTT, supplemented with 5 μM ZnSO 4 (top panels) or 5 mM EDTA (bottom panels). ‘D’ and ‘M’ indicate the elution volume corresponding to the size of a dimer (150 kDa) or a monomer (75 kDa), respectively ( B ) Gel filtration assays in buffer supplemented with 5 μM ZnSO 4 of WT, double mutant T1-SY (Cys308Ser and His359Tyr) and triple mutant T1-SYY (containing the additional mutation His488Tyr). Pictures on the left are based on the E. coli ThrRS structure (PDB code 1QF6) and represent the zinc-binding pocket showing in green the zinc coordination residues and in red residues mutated in the SY and SYY proteins. ( C ) Diagram showing the position of insertions b and c of T1 and insertion a of T2.

    Journal: Nucleic Acids Research

    Article Title: Trans-oligomerization of duplicated aminoacyl-tRNA synthetases maintains genetic code fidelity under stress

    doi: 10.1093/nar/gkv1020

    Figure Lengend Snippet: Cofactor loss promotes dissociation of T1. ( A ) Gel filtration assays of recombinant T1 and T2 proteins in buffer containing 150 mM NaCl, 100 mM Tris-HCl pH 7.5, 1 mM DTT, supplemented with 5 μM ZnSO 4 (top panels) or 5 mM EDTA (bottom panels). ‘D’ and ‘M’ indicate the elution volume corresponding to the size of a dimer (150 kDa) or a monomer (75 kDa), respectively ( B ) Gel filtration assays in buffer supplemented with 5 μM ZnSO 4 of WT, double mutant T1-SY (Cys308Ser and His359Tyr) and triple mutant T1-SYY (containing the additional mutation His488Tyr). Pictures on the left are based on the E. coli ThrRS structure (PDB code 1QF6) and represent the zinc-binding pocket showing in green the zinc coordination residues and in red residues mutated in the SY and SYY proteins. ( C ) Diagram showing the position of insertions b and c of T1 and insertion a of T2. "M1–3" indicate the conserved motifs of the catalytic domain of class II aaRSs. ( D ) Gel filtration of wild-type T1 or mutant proteins containing the indicated mutations using chromatography buffer supplemented with 5 mM EDTA.

    Article Snippet: For purification of His-tagged proteins by Ni-NTA chromatography, cell extracts were prepared in 100 mM Tris-HCl pH 8, 150 mM NaCl, 10% glycerol or 20 mM sodium phosphate pH 7.4, 500 mM NaCl and applied to His-Trap columns (GE Healthcare).

    Techniques: Filtration, Recombinant, Mutagenesis, Binding Assay, Chromatography

    In vitro replacement of the metal cofactor of T2. Pure preparations of recombinant T2 protein at concentration 25 μM were incubated with 5 mM EDTA and subsequently subjected to gel filtration in a Sephadex S-200 column equilibrated with Chelex 100-treated buffer containing 150 mM NaCl, 100 mM Tris-HCl pH 7.5, 1 mM DTT, supplemented with the metal indicated in each panel at a concentration of 5 μM. Aminoacylation activity was determined in fractions that contained protein and it is represented as gray bars. Fractions corresponding to the peak were subjected to buffer exchange to eliminate metals from the buffer and analyzed by ICP–MS. Numbers indicate the metal:protein ratio of the peak fractions.

    Journal: Nucleic Acids Research

    Article Title: Trans-oligomerization of duplicated aminoacyl-tRNA synthetases maintains genetic code fidelity under stress

    doi: 10.1093/nar/gkv1020

    Figure Lengend Snippet: In vitro replacement of the metal cofactor of T2. Pure preparations of recombinant T2 protein at concentration 25 μM were incubated with 5 mM EDTA and subsequently subjected to gel filtration in a Sephadex S-200 column equilibrated with Chelex 100-treated buffer containing 150 mM NaCl, 100 mM Tris-HCl pH 7.5, 1 mM DTT, supplemented with the metal indicated in each panel at a concentration of 5 μM. Aminoacylation activity was determined in fractions that contained protein and it is represented as gray bars. Fractions corresponding to the peak were subjected to buffer exchange to eliminate metals from the buffer and analyzed by ICP–MS. Numbers indicate the metal:protein ratio of the peak fractions.

    Article Snippet: For purification of His-tagged proteins by Ni-NTA chromatography, cell extracts were prepared in 100 mM Tris-HCl pH 8, 150 mM NaCl, 10% glycerol or 20 mM sodium phosphate pH 7.4, 500 mM NaCl and applied to His-Trap columns (GE Healthcare).

    Techniques: In Vitro, Recombinant, Concentration Assay, Incubation, Filtration, Activity Assay, Buffer Exchange, Mass Spectrometry

    Evaluation of contribution of YEM medium components for silver nanoparticles generation upon autoclaving with AgNO 3 . a : Alteration in color of 0.25 mM AgNO 3 upon autoclaving with different components of YEM medium namely, yeast extract (YE), mannitol, MgSO4, NaCl and K2HPO4 medium individually. b : UV-Vis absorption spectra of 0.25 mM AgNO 3 after autoclaving with YEM medium, yeast extract, mannitol, MgSO4, NaCl and K2HPO4.

    Journal: PLoS ONE

    Article Title: Inbuilt Potential of YEM Medium and Its Constituents to Generate Ag/Ag2O Nanoparticles

    doi: 10.1371/journal.pone.0061750

    Figure Lengend Snippet: Evaluation of contribution of YEM medium components for silver nanoparticles generation upon autoclaving with AgNO 3 . a : Alteration in color of 0.25 mM AgNO 3 upon autoclaving with different components of YEM medium namely, yeast extract (YE), mannitol, MgSO4, NaCl and K2HPO4 medium individually. b : UV-Vis absorption spectra of 0.25 mM AgNO 3 after autoclaving with YEM medium, yeast extract, mannitol, MgSO4, NaCl and K2HPO4.

    Article Snippet: Materials and Methods Mannitol, Yeast Extract, K2 HPO4 , MgSO4 .7H2 O and NaCl were of Himedia (India) make.

    Techniques: