hplc analysis Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher hplc analysis
    Daily main <t>SCFA</t> concentrations in fermentation effluents of IR and CR measured by <t>HPLC.</t> Initial stabilization: stabilization period in continuous mode to reach pseudo steady-state; closed symbol: IR; open symbol: CR; (▴, Δ) total SCFA, (•, ○) acetate, (♦, ◊) propionate, and (▪, □) butyrate.
    Hplc Analysis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1755 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Thermo Fisher
    Average 99 stars, based on 1755 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    99/100 stars
      Buy from Supplier

    99
    Millipore hplc analysis
    Daily main <t>SCFA</t> concentrations in fermentation effluents of IR and CR measured by <t>HPLC.</t> Initial stabilization: stabilization period in continuous mode to reach pseudo steady-state; closed symbol: IR; open symbol: CR; (▴, Δ) total SCFA, (•, ○) acetate, (♦, ◊) propionate, and (▪, □) butyrate.
    Hplc Analysis, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2012 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Millipore
    Average 99 stars, based on 2012 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    99/100 stars
      Buy from Supplier

    92
    Agilent technologies hplc analysis
    Overall structures of LmRPH at different states. ( A ) Structure of wild-type LmRPH in complex with <t>RIF,</t> ANP, and Mg 2+ . The AD, RD, and HD are colored lemon, light blue, and orange, respectively. ANP and RIF are shown as sticks and are colored green and magenta, respectively. Mg 2+ is shown as a sphere. ( B ) Gel-filtration profiles of wild-type LmRPH (red) and the G527A (green), G527S (cyan), and G527Y (blue) mutants. ( C ) In vitro catalytic activity of Gly527 mutants detected by <t>HPLC.</t> The reaction time of G527A, G527S, and G527Y is 1 h, and that of wild-type LmRPH is 5 min. Proteins were used at 0.5 mg/mL. ( D ) E . coli growth assay for Gly527 mutants. Bacteria transformed with wild-type LmRPH, vector, G527A, G527S, or G527Y were cultured in solid LB medium complemented with 0, 10, 40, or 80 μg/mL RIF. ( E ) Structure of LmRPH G527Y in apo form. ( F ) Structure of LmRPH G527Y in complex with ANP and Mg 2+ . Color codes in E and F are as A .
    Hplc Analysis, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 6036 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Agilent technologies
    Average 92 stars, based on 6036 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    92/100 stars
      Buy from Supplier

    94
    Waters Corporation hplc analysis
    A chromatogram of standard <t>ginsenosides</t> using <t>HPLC</t> assay. The column configuration consisted of an IMtakt Cadenza CD-C18 (4.6×75 mm). The UV absorption was measured at 203 nm. Gradient elution was employed, using solvent A (10% acetonitrile) and solvent B (90% acetonitrile) at 40℃. CK, compound K.
    Hplc Analysis, supplied by Waters Corporation, used in various techniques. Bioz Stars score: 94/100, based on 3928 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Waters Corporation
    Average 94 stars, based on 3928 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    94/100 stars
      Buy from Supplier

    94
    Shimadzu Corporation hplc analysis
    Average standard calibration curve ( n = 3) for urinary <t>rebamipide</t> determination at (A) 50–10,000 ng/mL and (B) 10,000–50,000 ng/mL. Each point represents urinary rebamipide detection under fluorescence <t>HPLC</t> set at 320 excitation and 380 emission. Also shown are the slope and y -intercept values used for determination of sample concentrations.
    Hplc Analysis, supplied by Shimadzu Corporation, used in various techniques. Bioz Stars score: 94/100, based on 3254 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Shimadzu Corporation
    Average 94 stars, based on 3254 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    94/100 stars
      Buy from Supplier

    94
    Phenomenex hplc analysis
    Synthesis of feruloylspermidine conjugates via recombinant Solanum melongena and S . richardii spermidine:hydroxycinnamoyl-CoA N -acyltansferase 1 orthologs (SmSHT and SrSHT). His-tagged recombinant SmSHT and SrSHT proteins were expressed in E . coli and purified from cell lysates by nickel affinity chromatography. ( a ) Evaluation of protein purity was performed via SDS-PAGE stained with Coomassie Brilliant Blue (left), and western blotting using anti-His-tag antibody (right): T, total cell lysate; P, cell pellet; S, supernatant; F, Flow through solution; W, last column wash; E1, E2 and E3, respectively, first, second and third column elutes. ( b ) <t>C18-HPLC-DAD</t> chromatogram of reaction products of SmSHT with spermidine plus feruloyl-CoA as substrates (mAU, milli-absorption units). ( c ) UV absorbance spectra of mono- (peaks 1a–1c), bis- (peaks 2a and 2b), and tris- (peak 3) N -feruloylspermidine conjugates.
    Hplc Analysis, supplied by Phenomenex, used in various techniques. Bioz Stars score: 94/100, based on 922 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Phenomenex
    Average 94 stars, based on 922 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    94/100 stars
      Buy from Supplier

    99
    Millipore hplc grade
    Synthesis of feruloylspermidine conjugates via recombinant Solanum melongena and S . richardii spermidine:hydroxycinnamoyl-CoA N -acyltansferase 1 orthologs (SmSHT and SrSHT). His-tagged recombinant SmSHT and SrSHT proteins were expressed in E . coli and purified from cell lysates by nickel affinity chromatography. ( a ) Evaluation of protein purity was performed via SDS-PAGE stained with Coomassie Brilliant Blue (left), and western blotting using anti-His-tag antibody (right): T, total cell lysate; P, cell pellet; S, supernatant; F, Flow through solution; W, last column wash; E1, E2 and E3, respectively, first, second and third column elutes. ( b ) <t>C18-HPLC-DAD</t> chromatogram of reaction products of SmSHT with spermidine plus feruloyl-CoA as substrates (mAU, milli-absorption units). ( c ) UV absorbance spectra of mono- (peaks 1a–1c), bis- (peaks 2a and 2b), and tris- (peak 3) N -feruloylspermidine conjugates.
    Hplc Grade, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2436 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc grade/product/Millipore
    Average 99 stars, based on 2436 article reviews
    Price from $9.99 to $1999.99
    hplc grade - by Bioz Stars, 2020-10
    99/100 stars
      Buy from Supplier

    99
    JASCO Inc hplc analysis
    Synthesis of feruloylspermidine conjugates via recombinant Solanum melongena and S . richardii spermidine:hydroxycinnamoyl-CoA N -acyltansferase 1 orthologs (SmSHT and SrSHT). His-tagged recombinant SmSHT and SrSHT proteins were expressed in E . coli and purified from cell lysates by nickel affinity chromatography. ( a ) Evaluation of protein purity was performed via SDS-PAGE stained with Coomassie Brilliant Blue (left), and western blotting using anti-His-tag antibody (right): T, total cell lysate; P, cell pellet; S, supernatant; F, Flow through solution; W, last column wash; E1, E2 and E3, respectively, first, second and third column elutes. ( b ) <t>C18-HPLC-DAD</t> chromatogram of reaction products of SmSHT with spermidine plus feruloyl-CoA as substrates (mAU, milli-absorption units). ( c ) UV absorbance spectra of mono- (peaks 1a–1c), bis- (peaks 2a and 2b), and tris- (peak 3) N -feruloylspermidine conjugates.
    Hplc Analysis, supplied by JASCO Inc, used in various techniques. Bioz Stars score: 99/100, based on 690 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/JASCO Inc
    Average 99 stars, based on 690 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    99/100 stars
      Buy from Supplier

    94
    Agilent technologies high performance liquid chromatography hplc analysis
    H 2 S deficiency increased the expression and activity of <t>IDO1</t> in Cse −/− mice. a <t>HPLC</t> analysis of Kyn/Trp ratios in wild-type (wt) mice and Cse −/− mice, N = 6. b , c Analysis of IDO1 expression in different tissues of wt mice and Cse −/− mice. Total RNA and protein were extracted from different tissues of wt mice and Cse −/− mice. mRNA expression of Ido1 was analyzed by qPCR, values from three independent experiments are presented as the mean ± SEM, * p
    High Performance Liquid Chromatography Hplc Analysis, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 94/100, based on 540 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/high performance liquid chromatography hplc analysis/product/Agilent technologies
    Average 94 stars, based on 540 article reviews
    Price from $9.99 to $1999.99
    high performance liquid chromatography hplc analysis - by Bioz Stars, 2020-10
    94/100 stars
      Buy from Supplier

    93
    Hitachi Ltd hplc analysis
    Degradation of 20 μM HHQ (A) and <t>PQS</t> (B) by recombinant E. coli strains. Cells suspended in LB medium were induced with 0.5 mM IPTG and incubated at 30°C overnight. Then they were supplemented with 20 μM HHQ or PQS and incubated at 30°C. Extracts of culture samples were analyzed by <t>HPLC.</t> (A) HHQ conversion by E. coli Rosetta pET28b:: mbp-aqdB , squares: HHQ, circles: PQS. (B) PQS conversion by E. coli BL21 pET28b:: mbp-aqdC ; circles: PQS, triangles: N -octanoylanthranilic acid (NOA). E. coli Rosetta and E. coli BL21 did not convert HHQ (discrete squares in A ) and PQS (discrete circles in B ).
    Hplc Analysis, supplied by Hitachi Ltd, used in various techniques. Bioz Stars score: 93/100, based on 487 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Hitachi Ltd
    Average 93 stars, based on 487 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    93/100 stars
      Buy from Supplier

    93
    Hewlett-Packard hplc analysis
    Identification of DIR genes. (A) ISP-1 resistance profiling of the DIR genes. Kinase-knockout ( DIR ) cells and control BY4741 (WT) cells were assessed for ISP-1 resistance on YPD-ISP-1 (750 ng mL −1 ) plates. The yeast culture was diluted serially as indicated and incubated at 30°C for 3 days. Plate images were acquired using the Printgraph camera system (ATTO, Tokyo, Japan). (B) LCB profiles of DIR cells. The LCB profiles of the DIR strains were determined in <t>HPLC-based</t> assays, as described in Experimental Procedures. The cellular levels of <t>C18-PHS</t> (white box) and C18-DHS (gray box) are expressed as incremented columns for each strain cultured in YPD medium (top). The LCB levels after 3 h of treatment with 500 ng mL −1 ISP-1 are also plotted (bottom). Each value shown is the mean of three independent experiments, and the standard deviation of error (SDE) for each experiment is indicated as a bar. Statistically significance differences compared to the WT control were evaluated using Student's t test and are shown when P
    Hplc Analysis, supplied by Hewlett-Packard, used in various techniques. Bioz Stars score: 93/100, based on 408 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Hewlett-Packard
    Average 93 stars, based on 408 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    93/100 stars
      Buy from Supplier

    92
    Shimadzu Corporation high performance liquid chromatography hplc analysis
    Three-dimensional chromatogram of <t>GMSYS</t> by <t>HPLC-PDA.</t> The retention times of 11 marker compounds—gallic acid, neomangiferin, chlorogenic acid, mangiferin, geniposide, paeoniflorin, berberine, liquiritin, nodakenin, glycyrrhizin, and atractylenolide III—were approximately 6.32, 10.82, 12.63, 13.04, 13.53, 15.17, 15.35, 16.61, 17.27, 29.86, and 34.00 min, respectively
    High Performance Liquid Chromatography Hplc Analysis, supplied by Shimadzu Corporation, used in various techniques. Bioz Stars score: 92/100, based on 322 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/high performance liquid chromatography hplc analysis/product/Shimadzu Corporation
    Average 92 stars, based on 322 article reviews
    Price from $9.99 to $1999.99
    high performance liquid chromatography hplc analysis - by Bioz Stars, 2020-10
    92/100 stars
      Buy from Supplier

    94
    Waters Corporation high performance liquid chromatography hplc analysis
    <t>HPLC</t> chromatogram for <t>DAE</t> (A), DHE (B) standard Betulinic acid (C). Arrow indicates peak for betulinic acid.
    High Performance Liquid Chromatography Hplc Analysis, supplied by Waters Corporation, used in various techniques. Bioz Stars score: 94/100, based on 411 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/high performance liquid chromatography hplc analysis/product/Waters Corporation
    Average 94 stars, based on 411 article reviews
    Price from $9.99 to $1999.99
    high performance liquid chromatography hplc analysis - by Bioz Stars, 2020-10
    94/100 stars
      Buy from Supplier

    92
    Bio-Rad hplc analysis
    Sections of chromatograms (between retention times 2.5 and 6.5 min) of a cocktail of known metabolites ( a , mix1), untreated CE population cell lysate ( b , 3P), cell lysate from CE populations exposed to 250 ppm ( c , 3L2), 500 ppm ( d , 3L5), 1000 ppm ( e , 3L10) and 2000 ppm ( f , 3L20) lead acetate, obtained by <t>CoulArray</t> <t>HPLC,</t> using the CoulArray software. a Chromatogram of a cocktail of known metabolites (1 mg/ml each). Uric acid eluted at 3.60 min (min) with its primary channel being 5; 8-hydroxy guanine at 3.98 min, channels 5 and 6; xanthine (xan) at 4.28 min, channels 12 and 13; guanine at 4.87 min, channels 11 and 12; guanosine at 5.28 min, channels 14 and 15; xanthosine at 5.56 min, channels 14 and 15; and, tyrosine at 5.93 min, channels 11, 12 and 13. b Chromatogram of sample 3PA [untreated 14-day CE population (P), the third replicate of set A , normalization factor 0.26]. c Chromatogram of sample 3L2A [14-day 250 ppm lead acetate treated CE population (L2), the third replicate of set A , normalization factor 0.41]. d Chromatogram of sample 3L5A [14-day 500 ppm lead acetate treated CE population (L5), the third replicate of set A , normalization factor 0.54]. e Chromatogram of sample 3L10A [14-day 1000 ppm lead acetate treated CE population (L10), the third replicate of set A, normalization factor 0.63]. f Chromatogram of sample 3L20A [14-day 2000 ppm lead acetate treated CE population (L20), the third replicate of set A, normalization factor 0.68]. One can see differences when comparing the chromatograms from the untreated to the various levels of lead treated nematode populations
    Hplc Analysis, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 257 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Bio-Rad
    Average 92 stars, based on 257 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    92/100 stars
      Buy from Supplier

    94
    Merck KGaA hplc analysis
    Evaluation of the intracellular <t>shikonin</t> concentration in shikonin-treated glioma cells. U87MG cells were treated with 2, 8, 50 and 100 μM of shikonin for 2 h. After treatment, cells were shattered by sonication and then centrifuged at 12,000 g for 10 min at 4°C. The cytosolic substances (supernatant) were evaluated the shikonin concentration detected by <t>HPLC.</t>
    Hplc Analysis, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 94/100, based on 279 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Merck KGaA
    Average 94 stars, based on 279 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    94/100 stars
      Buy from Supplier

    93
    Agilent technologies hplc ms ms analysis
    Effect of <t>HMNQ</t> and HQNO on the primary metabolome of E. coli . (A) Shown is the fold-change in the levels of 93 primary metabolites as a function of HMNQ or HQNO treatment, with respect to a no-treatment control, determined by <t>HR-HPLC-MS.</t> Three independent biological replicates for both drug-treated and untreated cells were used. The metabolites are sorted by fold-change. The color-coding is as follows: pyrimidine biosynthetic intermediates, red; NMPs, green; canonical amino acids, purple; NTPs, blue. (B) Biosynthetic pathway for pyrimidine nucleotides. HMNQ and HQNO inhibit DHODH, which results in accumulation of DHO and N-carbamoyl-L-aspartate (carbamoyl-Asp).
    Hplc Ms Ms Analysis, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 93/100, based on 552 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc ms ms analysis/product/Agilent technologies
    Average 93 stars, based on 552 article reviews
    Price from $9.99 to $1999.99
    hplc ms ms analysis - by Bioz Stars, 2020-10
    93/100 stars
      Buy from Supplier

    92
    Merck & Co hplc analysis
    (a and b) Calibration plot of the khellin using different concentrations versus peak area for <t>HPLC</t> and <t>HPTLC</t>
    Hplc Analysis, supplied by Merck & Co, used in various techniques. Bioz Stars score: 92/100, based on 145 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Merck & Co
    Average 92 stars, based on 145 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    92/100 stars
      Buy from Supplier

    92
    Gilson Inc hplc analysis
    <t>HPLC</t> chromatograms of SAT I (A) and SAT IV (B) assays with calf spleen microsomes; calf spleen microsomes were extracted with Triton. The resulting extract was tested for SAT I (A) and SAT IV (B) activity by incubation with the respective <t>BODIPY-labelled</t> substrates for 1 h at 37°C. The presence of the respective activity was indicated by the presence of the product (peak 2) next to unconverted substrate (peak 1).
    Hplc Analysis, supplied by Gilson Inc, used in various techniques. Bioz Stars score: 92/100, based on 264 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Gilson Inc
    Average 92 stars, based on 264 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    92/100 stars
      Buy from Supplier

    93
    Chromsystems Instruments hplc analysis
    Role of PGF 2α in adrenal endocrine functions. A , ELISA quantification of PGF 2α release by chromaffin <t>MPC862L</t> cells untreated (control) or treated with 10 −6 M dexamethasone for 12 h (dex). B , <t>HPLC</t> quantification of dopamine secretion by MPC862L cells cultured in absence or presence of 10 −7 M cloprostenol (clo) (PGF 2α analogue) used either alone or in combination with 10 −6 M dexamethasone for 12 h (clo+dex). Values are the mean of 3 experiments ± S.D. *, P
    Hplc Analysis, supplied by Chromsystems Instruments, used in various techniques. Bioz Stars score: 93/100, based on 130 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hplc analysis/product/Chromsystems Instruments
    Average 93 stars, based on 130 article reviews
    Price from $9.99 to $1999.99
    hplc analysis - by Bioz Stars, 2020-10
    93/100 stars
      Buy from Supplier

    Image Search Results


    Daily main SCFA concentrations in fermentation effluents of IR and CR measured by HPLC. Initial stabilization: stabilization period in continuous mode to reach pseudo steady-state; closed symbol: IR; open symbol: CR; (▴, Δ) total SCFA, (•, ○) acetate, (♦, ◊) propionate, and (▪, □) butyrate.

    Journal: PLoS ONE

    Article Title: In Vitro Continuous Fermentation Model (PolyFermS) of the Swine Proximal Colon for Simultaneous Testing on the Same Gut Microbiota

    doi: 10.1371/journal.pone.0094123

    Figure Lengend Snippet: Daily main SCFA concentrations in fermentation effluents of IR and CR measured by HPLC. Initial stabilization: stabilization period in continuous mode to reach pseudo steady-state; closed symbol: IR; open symbol: CR; (▴, Δ) total SCFA, (•, ○) acetate, (♦, ◊) propionate, and (▪, □) butyrate.

    Article Snippet: HPLC analysis for metabolite determination Short-chain fatty acids (SCFA; acetate, propionate, butyrate, valerate, formate, iso- butyrate and iso -valerate) as well as lactate concentrations in fermentation effluent samples from all reactors were determined by HPLC analysis (Thermo Fisher Scientific Inc. Accela, Wohlen, Switzerland).

    Techniques: High Performance Liquid Chromatography

    Overall structures of LmRPH at different states. ( A ) Structure of wild-type LmRPH in complex with RIF, ANP, and Mg 2+ . The AD, RD, and HD are colored lemon, light blue, and orange, respectively. ANP and RIF are shown as sticks and are colored green and magenta, respectively. Mg 2+ is shown as a sphere. ( B ) Gel-filtration profiles of wild-type LmRPH (red) and the G527A (green), G527S (cyan), and G527Y (blue) mutants. ( C ) In vitro catalytic activity of Gly527 mutants detected by HPLC. The reaction time of G527A, G527S, and G527Y is 1 h, and that of wild-type LmRPH is 5 min. Proteins were used at 0.5 mg/mL. ( D ) E . coli growth assay for Gly527 mutants. Bacteria transformed with wild-type LmRPH, vector, G527A, G527S, or G527Y were cultured in solid LB medium complemented with 0, 10, 40, or 80 μg/mL RIF. ( E ) Structure of LmRPH G527Y in apo form. ( F ) Structure of LmRPH G527Y in complex with ANP and Mg 2+ . Color codes in E and F are as A .

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Structural basis of rifampin inactivation by rifampin phosphotransferase

    doi: 10.1073/pnas.1523614113

    Figure Lengend Snippet: Overall structures of LmRPH at different states. ( A ) Structure of wild-type LmRPH in complex with RIF, ANP, and Mg 2+ . The AD, RD, and HD are colored lemon, light blue, and orange, respectively. ANP and RIF are shown as sticks and are colored green and magenta, respectively. Mg 2+ is shown as a sphere. ( B ) Gel-filtration profiles of wild-type LmRPH (red) and the G527A (green), G527S (cyan), and G527Y (blue) mutants. ( C ) In vitro catalytic activity of Gly527 mutants detected by HPLC. The reaction time of G527A, G527S, and G527Y is 1 h, and that of wild-type LmRPH is 5 min. Proteins were used at 0.5 mg/mL. ( D ) E . coli growth assay for Gly527 mutants. Bacteria transformed with wild-type LmRPH, vector, G527A, G527S, or G527Y were cultured in solid LB medium complemented with 0, 10, 40, or 80 μg/mL RIF. ( E ) Structure of LmRPH G527Y in apo form. ( F ) Structure of LmRPH G527Y in complex with ANP and Mg 2+ . Color codes in E and F are as A .

    Article Snippet: To identify the product RIF-P, the C18 column used for HPLC analysis was connected with an Agilent G6520A accurate-mass quadrupole time-of-flight LC-MS system (LC1200/MS Q-TOF6520).

    Techniques: Aqueous Normal-phase Chromatography, Filtration, In Vitro, Activity Assay, High Performance Liquid Chromatography, Growth Assay, Transformation Assay, Plasmid Preparation, Cell Culture

    Activity of LmRPH. ( A ) In vitro LmRPH reaction products analyzed by HPLC with the RIF detection program. ( B ) Identification of the two peaks in A by LC-MS. ( Left ) Peak 1. ( Right ) Peak 2. ( C ) The samples in A were analyzed by HPLC with a nucleotide-detection program. ( D ) E . coli growth assay. Bacteria transformed with LmRPH or vector were cultured in solid LB medium complemented with 0, 10, 100, or 1,000 μg/mL RIF.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Structural basis of rifampin inactivation by rifampin phosphotransferase

    doi: 10.1073/pnas.1523614113

    Figure Lengend Snippet: Activity of LmRPH. ( A ) In vitro LmRPH reaction products analyzed by HPLC with the RIF detection program. ( B ) Identification of the two peaks in A by LC-MS. ( Left ) Peak 1. ( Right ) Peak 2. ( C ) The samples in A were analyzed by HPLC with a nucleotide-detection program. ( D ) E . coli growth assay. Bacteria transformed with LmRPH or vector were cultured in solid LB medium complemented with 0, 10, 100, or 1,000 μg/mL RIF.

    Article Snippet: To identify the product RIF-P, the C18 column used for HPLC analysis was connected with an Agilent G6520A accurate-mass quadrupole time-of-flight LC-MS system (LC1200/MS Q-TOF6520).

    Techniques: Activity Assay, In Vitro, High Performance Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Growth Assay, Transformation Assay, Plasmid Preparation, Cell Culture

    Catalytic center of RIF phosphorylation. ( A ) Catalytic site of RIF phosphorylation. Residues His825, Lys670, Arg666, and Gln337 and RIF are shown as sticks; the water molecule is shown as a red sphere. Distances between the water molecule and surrounding residues are shown as dashed lines. ( B ) Catalytic site with a modeled RIF-P. Distances between the phosphate group and residues are shown as dashed lines. ( C ) In vitro catalytic activity of H825A, K670A, R666A, and Q337A detected by HPLC. ( D ) E . coli growth assay of the mutants in C. Bacteria transformed with wild-type LmRPH, vector, or mutants were cultured in solid LB medium complemented with 0 or 10 μg/mL RIF. ( E ) Phosphorylation analysis of wild-type LmRPH and the H825A mutant using Phos-tag SDS PAGE. LmRPH-P, phosphorylated LmRPH.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Structural basis of rifampin inactivation by rifampin phosphotransferase

    doi: 10.1073/pnas.1523614113

    Figure Lengend Snippet: Catalytic center of RIF phosphorylation. ( A ) Catalytic site of RIF phosphorylation. Residues His825, Lys670, Arg666, and Gln337 and RIF are shown as sticks; the water molecule is shown as a red sphere. Distances between the water molecule and surrounding residues are shown as dashed lines. ( B ) Catalytic site with a modeled RIF-P. Distances between the phosphate group and residues are shown as dashed lines. ( C ) In vitro catalytic activity of H825A, K670A, R666A, and Q337A detected by HPLC. ( D ) E . coli growth assay of the mutants in C. Bacteria transformed with wild-type LmRPH, vector, or mutants were cultured in solid LB medium complemented with 0 or 10 μg/mL RIF. ( E ) Phosphorylation analysis of wild-type LmRPH and the H825A mutant using Phos-tag SDS PAGE. LmRPH-P, phosphorylated LmRPH.

    Article Snippet: To identify the product RIF-P, the C18 column used for HPLC analysis was connected with an Agilent G6520A accurate-mass quadrupole time-of-flight LC-MS system (LC1200/MS Q-TOF6520).

    Techniques: In Vitro, Activity Assay, High Performance Liquid Chromatography, Growth Assay, Transformation Assay, Plasmid Preparation, Cell Culture, Mutagenesis, SDS Page

    ATP-binding site. ( A ) ATP-binding affinity of the AD (green isotherm), wild-type LmRPH (blue isotherm), and the G527Y mutant (red isotherm) measured by ITC. ( B ) Conformational changes of the AD induced by HD binding. The LmRPH–AD structure (gray) is superposed with the AD of the LmRPH G527Y –apo structure (lemon). The L13 loop connecting subdomains I and II is highlighted in red. The interaction interfaces between the AD and HD (orange) are shown in zoom-in views, and residues constituting the interface are shown with side chains. ( C ) Conformational changes induced in the AD and HD by ANP binding. The AD (lemon) and HD (orange) of the LmRPH G527Y –apo structure are superposed with those of the LmRPH G527Y -ANP structure (light blue). Structural elements undergoing conformational changes after ANP binding are colored in red. ( D ) Residues constituting the ATP-binding site. ANP (green) and residues (lemon) are shown as sticks, and Mg 2+ is shown as a lemon sphere. Coordination and hydrogen bonds are shown as dashed lines. ( E ) In vitro catalytic activity of ATP-binding site mutants detected by HPLC. The amounts of enzymes used in the assays of the K22A, R117A, E297A, T136A, Q309A, and R311A mutants are 10× those used in assays of wild-type LmRPH. ( F ) E . coli growth assay for ATP-binding site mutants. Bacteria transformed with wild-type LmRPH, vector, or mutants were cultured in solid LB medium complemented with 0, 10, 40, 80, or 320 μg/mL RIF.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Structural basis of rifampin inactivation by rifampin phosphotransferase

    doi: 10.1073/pnas.1523614113

    Figure Lengend Snippet: ATP-binding site. ( A ) ATP-binding affinity of the AD (green isotherm), wild-type LmRPH (blue isotherm), and the G527Y mutant (red isotherm) measured by ITC. ( B ) Conformational changes of the AD induced by HD binding. The LmRPH–AD structure (gray) is superposed with the AD of the LmRPH G527Y –apo structure (lemon). The L13 loop connecting subdomains I and II is highlighted in red. The interaction interfaces between the AD and HD (orange) are shown in zoom-in views, and residues constituting the interface are shown with side chains. ( C ) Conformational changes induced in the AD and HD by ANP binding. The AD (lemon) and HD (orange) of the LmRPH G527Y –apo structure are superposed with those of the LmRPH G527Y -ANP structure (light blue). Structural elements undergoing conformational changes after ANP binding are colored in red. ( D ) Residues constituting the ATP-binding site. ANP (green) and residues (lemon) are shown as sticks, and Mg 2+ is shown as a lemon sphere. Coordination and hydrogen bonds are shown as dashed lines. ( E ) In vitro catalytic activity of ATP-binding site mutants detected by HPLC. The amounts of enzymes used in the assays of the K22A, R117A, E297A, T136A, Q309A, and R311A mutants are 10× those used in assays of wild-type LmRPH. ( F ) E . coli growth assay for ATP-binding site mutants. Bacteria transformed with wild-type LmRPH, vector, or mutants were cultured in solid LB medium complemented with 0, 10, 40, 80, or 320 μg/mL RIF.

    Article Snippet: To identify the product RIF-P, the C18 column used for HPLC analysis was connected with an Agilent G6520A accurate-mass quadrupole time-of-flight LC-MS system (LC1200/MS Q-TOF6520).

    Techniques: Binding Assay, Mutagenesis, Aqueous Normal-phase Chromatography, In Vitro, Activity Assay, High Performance Liquid Chromatography, Growth Assay, Transformation Assay, Plasmid Preparation, Cell Culture

    A chromatogram of standard ginsenosides using HPLC assay. The column configuration consisted of an IMtakt Cadenza CD-C18 (4.6×75 mm). The UV absorption was measured at 203 nm. Gradient elution was employed, using solvent A (10% acetonitrile) and solvent B (90% acetonitrile) at 40℃. CK, compound K.

    Journal: Journal of Ginseng Research

    Article Title: The bioavailability of red ginseng extract fermented by Phellinus linteus

    doi: 10.5142/jgr.2013.37.108

    Figure Lengend Snippet: A chromatogram of standard ginsenosides using HPLC assay. The column configuration consisted of an IMtakt Cadenza CD-C18 (4.6×75 mm). The UV absorption was measured at 203 nm. Gradient elution was employed, using solvent A (10% acetonitrile) and solvent B (90% acetonitrile) at 40℃. CK, compound K.

    Article Snippet: HPLC analysis of ginsenosides A purified sample for HPLC analysis was prepared by C18 ODS cartridge (Waters Associates, Milford, MA, USA) solid-phase extraction according to a method described previously .

    Techniques: High Performance Liquid Chromatography

    A chromatogram of (A) non-fermented red ginseng and (B) fermented red ginseng by Phellinus linteus ginsenosides using HPLC assay. The column configuration consisted of an IMtakt Cadenza CD-C18 (4.6×75 mm). The UV absorption was measured at 203 nm. Gradient elution was employed, using solvent A (10% acetonitrile) and solvent B (90% acetonitrile) at 40℃.

    Journal: Journal of Ginseng Research

    Article Title: The bioavailability of red ginseng extract fermented by Phellinus linteus

    doi: 10.5142/jgr.2013.37.108

    Figure Lengend Snippet: A chromatogram of (A) non-fermented red ginseng and (B) fermented red ginseng by Phellinus linteus ginsenosides using HPLC assay. The column configuration consisted of an IMtakt Cadenza CD-C18 (4.6×75 mm). The UV absorption was measured at 203 nm. Gradient elution was employed, using solvent A (10% acetonitrile) and solvent B (90% acetonitrile) at 40℃.

    Article Snippet: HPLC analysis of ginsenosides A purified sample for HPLC analysis was prepared by C18 ODS cartridge (Waters Associates, Milford, MA, USA) solid-phase extraction according to a method described previously .

    Techniques: High Performance Liquid Chromatography

    Average standard calibration curve ( n = 3) for urinary rebamipide determination at (A) 50–10,000 ng/mL and (B) 10,000–50,000 ng/mL. Each point represents urinary rebamipide detection under fluorescence HPLC set at 320 excitation and 380 emission. Also shown are the slope and y -intercept values used for determination of sample concentrations.

    Journal: MethodsX

    Article Title: A simple high performance liquid chromatography method for determination of rebamipide in rat urine

    doi: 10.1016/j.mex.2014.06.002

    Figure Lengend Snippet: Average standard calibration curve ( n = 3) for urinary rebamipide determination at (A) 50–10,000 ng/mL and (B) 10,000–50,000 ng/mL. Each point represents urinary rebamipide detection under fluorescence HPLC set at 320 excitation and 380 emission. Also shown are the slope and y -intercept values used for determination of sample concentrations.

    Article Snippet: Analysis of rebamipide concentrations in the urine involved preparation of rebamipide stock solution, development of low and high linear standard calibration curves to compensate for altered rebamipide concentrations noted in urine samples, use of acetonitrile and hydrochloric acid for urinary rebamipide extraction, and HPLC analysis of rebamipide using a Shimadzu HPLC system.

    Techniques: Fluorescence, High Performance Liquid Chromatography

    Representative HPLC chromatogram of (A) rat urine spiked with 10,000 ng/mL rebamipide and (B) rat urine spiked with 5000 ng/mL rebamipide.

    Journal: MethodsX

    Article Title: A simple high performance liquid chromatography method for determination of rebamipide in rat urine

    doi: 10.1016/j.mex.2014.06.002

    Figure Lengend Snippet: Representative HPLC chromatogram of (A) rat urine spiked with 10,000 ng/mL rebamipide and (B) rat urine spiked with 5000 ng/mL rebamipide.

    Article Snippet: Analysis of rebamipide concentrations in the urine involved preparation of rebamipide stock solution, development of low and high linear standard calibration curves to compensate for altered rebamipide concentrations noted in urine samples, use of acetonitrile and hydrochloric acid for urinary rebamipide extraction, and HPLC analysis of rebamipide using a Shimadzu HPLC system.

    Techniques: High Performance Liquid Chromatography

    Representative HPLC chromatogram of urine sample obtained from a rat dosed with rebamipide.

    Journal: MethodsX

    Article Title: A simple high performance liquid chromatography method for determination of rebamipide in rat urine

    doi: 10.1016/j.mex.2014.06.002

    Figure Lengend Snippet: Representative HPLC chromatogram of urine sample obtained from a rat dosed with rebamipide.

    Article Snippet: Analysis of rebamipide concentrations in the urine involved preparation of rebamipide stock solution, development of low and high linear standard calibration curves to compensate for altered rebamipide concentrations noted in urine samples, use of acetonitrile and hydrochloric acid for urinary rebamipide extraction, and HPLC analysis of rebamipide using a Shimadzu HPLC system.

    Techniques: High Performance Liquid Chromatography

    Synthesis of feruloylspermidine conjugates via recombinant Solanum melongena and S . richardii spermidine:hydroxycinnamoyl-CoA N -acyltansferase 1 orthologs (SmSHT and SrSHT). His-tagged recombinant SmSHT and SrSHT proteins were expressed in E . coli and purified from cell lysates by nickel affinity chromatography. ( a ) Evaluation of protein purity was performed via SDS-PAGE stained with Coomassie Brilliant Blue (left), and western blotting using anti-His-tag antibody (right): T, total cell lysate; P, cell pellet; S, supernatant; F, Flow through solution; W, last column wash; E1, E2 and E3, respectively, first, second and third column elutes. ( b ) C18-HPLC-DAD chromatogram of reaction products of SmSHT with spermidine plus feruloyl-CoA as substrates (mAU, milli-absorption units). ( c ) UV absorbance spectra of mono- (peaks 1a–1c), bis- (peaks 2a and 2b), and tris- (peak 3) N -feruloylspermidine conjugates.

    Journal: Horticulture Research

    Article Title: Characterization of spermidine hydroxycinnamoyl transferases from eggplant (Solanum melongena L.) and its wild relative Solanum richardii Dunal

    doi: 10.1038/hortres.2016.62

    Figure Lengend Snippet: Synthesis of feruloylspermidine conjugates via recombinant Solanum melongena and S . richardii spermidine:hydroxycinnamoyl-CoA N -acyltansferase 1 orthologs (SmSHT and SrSHT). His-tagged recombinant SmSHT and SrSHT proteins were expressed in E . coli and purified from cell lysates by nickel affinity chromatography. ( a ) Evaluation of protein purity was performed via SDS-PAGE stained with Coomassie Brilliant Blue (left), and western blotting using anti-His-tag antibody (right): T, total cell lysate; P, cell pellet; S, supernatant; F, Flow through solution; W, last column wash; E1, E2 and E3, respectively, first, second and third column elutes. ( b ) C18-HPLC-DAD chromatogram of reaction products of SmSHT with spermidine plus feruloyl-CoA as substrates (mAU, milli-absorption units). ( c ) UV absorbance spectra of mono- (peaks 1a–1c), bis- (peaks 2a and 2b), and tris- (peak 3) N -feruloylspermidine conjugates.

    Article Snippet: HPLC-DAD analysis of hydroxycinnamoyl-CoAs and HCAAs For HPLC analysis of Sm4CL1 reaction, samples were fractionated on 1 g, 33 μm Strata X polymeric reverse phase, solid phase extraction (SPE) tubes (Phenomenex, Torrance, CA, USA) with a step gradient of pH 8 aqueous Ammonium Hydroxide, 80% aqueous MeOH and 100% MeOH.

    Techniques: Recombinant, Purification, Affinity Chromatography, SDS Page, Staining, Western Blot, Flow Cytometry, High Performance Liquid Chromatography

    Synthesis of caffeoyl-CoA via Solanum melongena 4-coumarate:CoA ligase 1 (Sm4CL1). His-tagged recombinant Sm4CL1 protein was expressed in E. coli and purified from cell lysates by nickel affinity chromatography. ( a ) Evaluation of protein purity was performed via SDS-PAGE stained with Coomassie Brilliant Blue (left), and Western blotting using anti-His-tag antibody (right): T, total cell lysate; P, cell pellet; S, supernatant; W, last column wash; E1, E2 and E3, respectively, first, second and third column elutes. ( b ), ( c ) and ( d ) in order: C18-HPLC-DAD chromatograms of a caffeic acid standard, the substrate plus reaction product of Sm4CL1, and purified caffeoyl-CoA from the Sm4CL1 reaction (mAU, milli-absorbance units). ( e ) UV absorbance spectra of caffeic acid (substrate) and caffeoyl-CoA (product).

    Journal: Horticulture Research

    Article Title: Characterization of spermidine hydroxycinnamoyl transferases from eggplant (Solanum melongena L.) and its wild relative Solanum richardii Dunal

    doi: 10.1038/hortres.2016.62

    Figure Lengend Snippet: Synthesis of caffeoyl-CoA via Solanum melongena 4-coumarate:CoA ligase 1 (Sm4CL1). His-tagged recombinant Sm4CL1 protein was expressed in E. coli and purified from cell lysates by nickel affinity chromatography. ( a ) Evaluation of protein purity was performed via SDS-PAGE stained with Coomassie Brilliant Blue (left), and Western blotting using anti-His-tag antibody (right): T, total cell lysate; P, cell pellet; S, supernatant; W, last column wash; E1, E2 and E3, respectively, first, second and third column elutes. ( b ), ( c ) and ( d ) in order: C18-HPLC-DAD chromatograms of a caffeic acid standard, the substrate plus reaction product of Sm4CL1, and purified caffeoyl-CoA from the Sm4CL1 reaction (mAU, milli-absorbance units). ( e ) UV absorbance spectra of caffeic acid (substrate) and caffeoyl-CoA (product).

    Article Snippet: HPLC-DAD analysis of hydroxycinnamoyl-CoAs and HCAAs For HPLC analysis of Sm4CL1 reaction, samples were fractionated on 1 g, 33 μm Strata X polymeric reverse phase, solid phase extraction (SPE) tubes (Phenomenex, Torrance, CA, USA) with a step gradient of pH 8 aqueous Ammonium Hydroxide, 80% aqueous MeOH and 100% MeOH.

    Techniques: Recombinant, Purification, Affinity Chromatography, SDS Page, Staining, Western Blot, High Performance Liquid Chromatography

    H 2 S deficiency increased the expression and activity of IDO1 in Cse −/− mice. a HPLC analysis of Kyn/Trp ratios in wild-type (wt) mice and Cse −/− mice, N = 6. b , c Analysis of IDO1 expression in different tissues of wt mice and Cse −/− mice. Total RNA and protein were extracted from different tissues of wt mice and Cse −/− mice. mRNA expression of Ido1 was analyzed by qPCR, values from three independent experiments are presented as the mean ± SEM, * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: H2S suppresses indoleamine 2, 3-dioxygenase 1 and exhibits immunotherapeutic efficacy in murine hepatocellular carcinoma

    doi: 10.1186/s13046-019-1083-5

    Figure Lengend Snippet: H 2 S deficiency increased the expression and activity of IDO1 in Cse −/− mice. a HPLC analysis of Kyn/Trp ratios in wild-type (wt) mice and Cse −/− mice, N = 6. b , c Analysis of IDO1 expression in different tissues of wt mice and Cse −/− mice. Total RNA and protein were extracted from different tissues of wt mice and Cse −/− mice. mRNA expression of Ido1 was analyzed by qPCR, values from three independent experiments are presented as the mean ± SEM, * p

    Article Snippet: High-performance liquid chromatography (HPLC) analysis The IDO1 activities of mouse serum and cell supernatant were analyzed by an Agilent 1260 series HPLC system (Agilent Corp., USA).

    Techniques: Expressing, Activity Assay, Mouse Assay, High Performance Liquid Chromatography, Real-time Polymerase Chain Reaction

    H 2 S inhibited IDO1 activity via H 2 S/NO crosstalk, and NO rather than H 2 S was an IDO1 inhibitor. a H 2 S donor upregulated the mRNA expression of iNOS in MCF-7 and SGC-7901 cells. Total RNA was extracted from the cells that were treated with different concentrations of NaHS for 24 h, and the mRNA expression levels of iNOS were analyzed by qPCR. b H 2 S deficiency downregulated the mRNA expression of Inos in Cse −/− mice. Total RNA was extracted from different tissues of the wt mice and Cse −/− mice, and the mRNA expression levels of Inos were analyzed by qPCR. c NO played an important role in the regulation of H 2 S on IDO1 activity. The cell supernatants were harvested, and kynurenine and tryptophan levels were measured by HPLC. d , e NO rather than H 2 S was a direct inhibitor of IDO1. Enzymatic assay of IDO1 was carried out with recombinant human IDO1 (rhIDO1) and different concentrations (0–400 μM) of H 2 S donors (NaHS, GYY4137), NO donor (SNP) and 1-L-MT. f Schematic diagram depicting the modulation of H 2 S on IDO1. All experiments repeated at least three times. Statistical significance was determined by Student’s t test ( b ) and one-way ANOVA followed by Dunnett’s test ( a , c and d ). * P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: H2S suppresses indoleamine 2, 3-dioxygenase 1 and exhibits immunotherapeutic efficacy in murine hepatocellular carcinoma

    doi: 10.1186/s13046-019-1083-5

    Figure Lengend Snippet: H 2 S inhibited IDO1 activity via H 2 S/NO crosstalk, and NO rather than H 2 S was an IDO1 inhibitor. a H 2 S donor upregulated the mRNA expression of iNOS in MCF-7 and SGC-7901 cells. Total RNA was extracted from the cells that were treated with different concentrations of NaHS for 24 h, and the mRNA expression levels of iNOS were analyzed by qPCR. b H 2 S deficiency downregulated the mRNA expression of Inos in Cse −/− mice. Total RNA was extracted from different tissues of the wt mice and Cse −/− mice, and the mRNA expression levels of Inos were analyzed by qPCR. c NO played an important role in the regulation of H 2 S on IDO1 activity. The cell supernatants were harvested, and kynurenine and tryptophan levels were measured by HPLC. d , e NO rather than H 2 S was a direct inhibitor of IDO1. Enzymatic assay of IDO1 was carried out with recombinant human IDO1 (rhIDO1) and different concentrations (0–400 μM) of H 2 S donors (NaHS, GYY4137), NO donor (SNP) and 1-L-MT. f Schematic diagram depicting the modulation of H 2 S on IDO1. All experiments repeated at least three times. Statistical significance was determined by Student’s t test ( b ) and one-way ANOVA followed by Dunnett’s test ( a , c and d ). * P

    Article Snippet: High-performance liquid chromatography (HPLC) analysis The IDO1 activities of mouse serum and cell supernatant were analyzed by an Agilent 1260 series HPLC system (Agilent Corp., USA).

    Techniques: Activity Assay, Expressing, Real-time Polymerase Chain Reaction, Mouse Assay, High Performance Liquid Chromatography, Enzymatic Assay, Recombinant

    Degradation of 20 μM HHQ (A) and PQS (B) by recombinant E. coli strains. Cells suspended in LB medium were induced with 0.5 mM IPTG and incubated at 30°C overnight. Then they were supplemented with 20 μM HHQ or PQS and incubated at 30°C. Extracts of culture samples were analyzed by HPLC. (A) HHQ conversion by E. coli Rosetta pET28b:: mbp-aqdB , squares: HHQ, circles: PQS. (B) PQS conversion by E. coli BL21 pET28b:: mbp-aqdC ; circles: PQS, triangles: N -octanoylanthranilic acid (NOA). E. coli Rosetta and E. coli BL21 did not convert HHQ (discrete squares in A ) and PQS (discrete circles in B ).

    Journal: Frontiers in Microbiology

    Article Title: Mycobacterium abscessus subsp. abscessus Is Capable of Degrading Pseudomonas aeruginosa Quinolone Signals

    doi: 10.3389/fmicb.2017.00339

    Figure Lengend Snippet: Degradation of 20 μM HHQ (A) and PQS (B) by recombinant E. coli strains. Cells suspended in LB medium were induced with 0.5 mM IPTG and incubated at 30°C overnight. Then they were supplemented with 20 μM HHQ or PQS and incubated at 30°C. Extracts of culture samples were analyzed by HPLC. (A) HHQ conversion by E. coli Rosetta pET28b:: mbp-aqdB , squares: HHQ, circles: PQS. (B) PQS conversion by E. coli BL21 pET28b:: mbp-aqdC ; circles: PQS, triangles: N -octanoylanthranilic acid (NOA). E. coli Rosetta and E. coli BL21 did not convert HHQ (discrete squares in A ) and PQS (discrete circles in B ).

    Article Snippet: HPLC Analysis For the identification and quantification of PQS and other AQs, compounds were separated on a 250 × 4 mm Eurospher II RP-18 column using a Hitachi EZchrom Elite HPLC system with diode array detector model 2450, or an Agilent 1100 series system with diode array detector model G1315B.

    Techniques: Recombinant, Incubation, High Performance Liquid Chromatography

    HHQ and PQS conversion by cell extracts of M. abscessus T . Extracts were obtained from cells grown in DSM219 medium (filled symbols), and from cells grown in the same medium but supplemented with PQS 2 h prior to harvesting (open symbols). Extracts obtained by sonication of cell suspensions were centrifuged, the resulting supernatants were desalted, set to a protein concentration of 1 mg/mL in 50 mM potassium phosphate buffer pH 7.5, and incubated with 20 μM HHQ and 500 μM NADH (A) , or with 20 μM PQS (B) . Samples were withdrawn at the given time points, extracted, and analyzed by HPLC. Means ± SD of three biological replicates are shown.

    Journal: Frontiers in Microbiology

    Article Title: Mycobacterium abscessus subsp. abscessus Is Capable of Degrading Pseudomonas aeruginosa Quinolone Signals

    doi: 10.3389/fmicb.2017.00339

    Figure Lengend Snippet: HHQ and PQS conversion by cell extracts of M. abscessus T . Extracts were obtained from cells grown in DSM219 medium (filled symbols), and from cells grown in the same medium but supplemented with PQS 2 h prior to harvesting (open symbols). Extracts obtained by sonication of cell suspensions were centrifuged, the resulting supernatants were desalted, set to a protein concentration of 1 mg/mL in 50 mM potassium phosphate buffer pH 7.5, and incubated with 20 μM HHQ and 500 μM NADH (A) , or with 20 μM PQS (B) . Samples were withdrawn at the given time points, extracted, and analyzed by HPLC. Means ± SD of three biological replicates are shown.

    Article Snippet: HPLC Analysis For the identification and quantification of PQS and other AQs, compounds were separated on a 250 × 4 mm Eurospher II RP-18 column using a Hitachi EZchrom Elite HPLC system with diode array detector model 2450, or an Agilent 1100 series system with diode array detector model G1315B.

    Techniques: Sonication, Protein Concentration, Incubation, High Performance Liquid Chromatography

    Degradation of 20 μM HHQ (A) and PQS (B) by M. abscessus strains. Cells suspended at an OD 600 nm of 3.5 in DSM219 medium were supplemented with 20 μM HHQ or PQS, and incubated at 37°C. Extracts of culture samples were analyzed by HPLC. Squares: M. abscessus P4a, circles: M. abscessus P13, triangles: M. abscessus P5a. Means ± SD of three biological replicates are shown.

    Journal: Frontiers in Microbiology

    Article Title: Mycobacterium abscessus subsp. abscessus Is Capable of Degrading Pseudomonas aeruginosa Quinolone Signals

    doi: 10.3389/fmicb.2017.00339

    Figure Lengend Snippet: Degradation of 20 μM HHQ (A) and PQS (B) by M. abscessus strains. Cells suspended at an OD 600 nm of 3.5 in DSM219 medium were supplemented with 20 μM HHQ or PQS, and incubated at 37°C. Extracts of culture samples were analyzed by HPLC. Squares: M. abscessus P4a, circles: M. abscessus P13, triangles: M. abscessus P5a. Means ± SD of three biological replicates are shown.

    Article Snippet: HPLC Analysis For the identification and quantification of PQS and other AQs, compounds were separated on a 250 × 4 mm Eurospher II RP-18 column using a Hitachi EZchrom Elite HPLC system with diode array detector model 2450, or an Agilent 1100 series system with diode array detector model G1315B.

    Techniques: Incubation, High Performance Liquid Chromatography

    Pseudomonas quinolone signal consumption by the type strain and clinical isolates of Mycobacterium abscessus . Cells suspended at an OD 600 nm of 3.5 in DSM219 medium were supplemented with 20 μM PQS and incubated at 37°C. Extracts of culture samples withdrawn after 0, 4 and 24 h were analyzed by HPLC. The PQS concentrations in samples taken at t = 0 h were set as 100%. Means ± SE of three independent biological replicates are shown. Decrease in PQS concentrations in sterile DSM219 medium due to abiotic oxidation was not observed within 24 h.

    Journal: Frontiers in Microbiology

    Article Title: Mycobacterium abscessus subsp. abscessus Is Capable of Degrading Pseudomonas aeruginosa Quinolone Signals

    doi: 10.3389/fmicb.2017.00339

    Figure Lengend Snippet: Pseudomonas quinolone signal consumption by the type strain and clinical isolates of Mycobacterium abscessus . Cells suspended at an OD 600 nm of 3.5 in DSM219 medium were supplemented with 20 μM PQS and incubated at 37°C. Extracts of culture samples withdrawn after 0, 4 and 24 h were analyzed by HPLC. The PQS concentrations in samples taken at t = 0 h were set as 100%. Means ± SE of three independent biological replicates are shown. Decrease in PQS concentrations in sterile DSM219 medium due to abiotic oxidation was not observed within 24 h.

    Article Snippet: HPLC Analysis For the identification and quantification of PQS and other AQs, compounds were separated on a 250 × 4 mm Eurospher II RP-18 column using a Hitachi EZchrom Elite HPLC system with diode array detector model 2450, or an Agilent 1100 series system with diode array detector model G1315B.

    Techniques: Incubation, High Performance Liquid Chromatography

    Relative PQS levels in cultures of P. aeruginosa PAO1 (white bars) and co-cultures of P. aeruginosa PAO1 with M. abscessus P4a (light gray), M. abscessus P5a (gray), M. abscessus P13 (dark gray), and autoclaved M. abscessus P13 (black), incubated under shaking at 37°C. Extracts of cultures were analyzed by HPLC. PQS concentrations in P. aeruginosa PAO1 cultures were set as 1 (8 h: 1.79 μM; 24 h: 0.76 μM PQS). Means ± SD of three biological replicates are shown. CFUs of P. aeruginosa PAO1 after 24 h of incubation: PAO1 culture: 1.81 × 10 11 ; PAO1 – P13 co-culture: 1.21 × 10 11 ; PAO1 cultured with autoclaved P13: 1.32 × 10 11 .

    Journal: Frontiers in Microbiology

    Article Title: Mycobacterium abscessus subsp. abscessus Is Capable of Degrading Pseudomonas aeruginosa Quinolone Signals

    doi: 10.3389/fmicb.2017.00339

    Figure Lengend Snippet: Relative PQS levels in cultures of P. aeruginosa PAO1 (white bars) and co-cultures of P. aeruginosa PAO1 with M. abscessus P4a (light gray), M. abscessus P5a (gray), M. abscessus P13 (dark gray), and autoclaved M. abscessus P13 (black), incubated under shaking at 37°C. Extracts of cultures were analyzed by HPLC. PQS concentrations in P. aeruginosa PAO1 cultures were set as 1 (8 h: 1.79 μM; 24 h: 0.76 μM PQS). Means ± SD of three biological replicates are shown. CFUs of P. aeruginosa PAO1 after 24 h of incubation: PAO1 culture: 1.81 × 10 11 ; PAO1 – P13 co-culture: 1.21 × 10 11 ; PAO1 cultured with autoclaved P13: 1.32 × 10 11 .

    Article Snippet: HPLC Analysis For the identification and quantification of PQS and other AQs, compounds were separated on a 250 × 4 mm Eurospher II RP-18 column using a Hitachi EZchrom Elite HPLC system with diode array detector model 2450, or an Agilent 1100 series system with diode array detector model G1315B.

    Techniques: Incubation, High Performance Liquid Chromatography, Co-Culture Assay, Cell Culture

    Identification of DIR genes. (A) ISP-1 resistance profiling of the DIR genes. Kinase-knockout ( DIR ) cells and control BY4741 (WT) cells were assessed for ISP-1 resistance on YPD-ISP-1 (750 ng mL −1 ) plates. The yeast culture was diluted serially as indicated and incubated at 30°C for 3 days. Plate images were acquired using the Printgraph camera system (ATTO, Tokyo, Japan). (B) LCB profiles of DIR cells. The LCB profiles of the DIR strains were determined in HPLC-based assays, as described in Experimental Procedures. The cellular levels of C18-PHS (white box) and C18-DHS (gray box) are expressed as incremented columns for each strain cultured in YPD medium (top). The LCB levels after 3 h of treatment with 500 ng mL −1 ISP-1 are also plotted (bottom). Each value shown is the mean of three independent experiments, and the standard deviation of error (SDE) for each experiment is indicated as a bar. Statistically significance differences compared to the WT control were evaluated using Student's t test and are shown when P

    Journal: MicrobiologyOpen

    Article Title: Fpk1/2 kinases regulate cellular sphingoid long-chain base abundance and alter cellular resistance to LCB elevation or depletion

    doi: 10.1002/mbo3.160

    Figure Lengend Snippet: Identification of DIR genes. (A) ISP-1 resistance profiling of the DIR genes. Kinase-knockout ( DIR ) cells and control BY4741 (WT) cells were assessed for ISP-1 resistance on YPD-ISP-1 (750 ng mL −1 ) plates. The yeast culture was diluted serially as indicated and incubated at 30°C for 3 days. Plate images were acquired using the Printgraph camera system (ATTO, Tokyo, Japan). (B) LCB profiles of DIR cells. The LCB profiles of the DIR strains were determined in HPLC-based assays, as described in Experimental Procedures. The cellular levels of C18-PHS (white box) and C18-DHS (gray box) are expressed as incremented columns for each strain cultured in YPD medium (top). The LCB levels after 3 h of treatment with 500 ng mL −1 ISP-1 are also plotted (bottom). Each value shown is the mean of three independent experiments, and the standard deviation of error (SDE) for each experiment is indicated as a bar. Statistically significance differences compared to the WT control were evaluated using Student's t test and are shown when P

    Article Snippet: Briefly, HPLC analysis was performed using a C18 column (4.6 × 250 mm, XDB-C18; Hewlett-Packard, Palo Alto, CA) on a Shimadzu LC10A series liquid chromatography system.

    Techniques: Knock-Out, Incubation, High Performance Liquid Chromatography, Cell Culture, Standard Deviation

    LCB levels are increased in the fpk1/2Δ mutant. (A) LCB levels in fpk1/2Δ cells. LCB levels were measured in the indicated mutant strains before and after ISP-1 treatment in YPD medium as in Figure 1 B. LCB levels were measured by HPLC, and the values in the columns indicate the mean levels of C18-PHS and C18-DHS with SDE values from three independent experiments. * P

    Journal: MicrobiologyOpen

    Article Title: Fpk1/2 kinases regulate cellular sphingoid long-chain base abundance and alter cellular resistance to LCB elevation or depletion

    doi: 10.1002/mbo3.160

    Figure Lengend Snippet: LCB levels are increased in the fpk1/2Δ mutant. (A) LCB levels in fpk1/2Δ cells. LCB levels were measured in the indicated mutant strains before and after ISP-1 treatment in YPD medium as in Figure 1 B. LCB levels were measured by HPLC, and the values in the columns indicate the mean levels of C18-PHS and C18-DHS with SDE values from three independent experiments. * P

    Article Snippet: Briefly, HPLC analysis was performed using a C18 column (4.6 × 250 mm, XDB-C18; Hewlett-Packard, Palo Alto, CA) on a Shimadzu LC10A series liquid chromatography system.

    Techniques: Mutagenesis, High Performance Liquid Chromatography

    Three-dimensional chromatogram of GMSYS by HPLC-PDA. The retention times of 11 marker compounds—gallic acid, neomangiferin, chlorogenic acid, mangiferin, geniposide, paeoniflorin, berberine, liquiritin, nodakenin, glycyrrhizin, and atractylenolide III—were approximately 6.32, 10.82, 12.63, 13.04, 13.53, 15.17, 15.35, 16.61, 17.27, 29.86, and 34.00 min, respectively

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Anti-inflammatory effect and action mechanisms of traditional herbal formula Gamisoyo-san in RAW 264.7 macrophages

    doi: 10.1186/s12906-016-1197-7

    Figure Lengend Snippet: Three-dimensional chromatogram of GMSYS by HPLC-PDA. The retention times of 11 marker compounds—gallic acid, neomangiferin, chlorogenic acid, mangiferin, geniposide, paeoniflorin, berberine, liquiritin, nodakenin, glycyrrhizin, and atractylenolide III—were approximately 6.32, 10.82, 12.63, 13.04, 13.53, 15.17, 15.35, 16.61, 17.27, 29.86, and 34.00 min, respectively

    Article Snippet: High-performance liquid chromatography (HPLC) analysis of GMSYS Quantitative analysis of the GMSYS sample was performed using an LC-20A Prominence HPLC system (Shimadzu Corp., Kyoto, Japan) equipped with a solvent delivery unit, an on-line degasser, a column oven, an autosampler, and a photo diode array (PDA) detector.

    Techniques: High Performance Liquid Chromatography, Marker

    HPLC chromatogram for DAE (A), DHE (B) standard Betulinic acid (C). Arrow indicates peak for betulinic acid.

    Journal: Journal of Traditional and Complementary Medicine

    Article Title: Dillenia indica L. attenuates diabetic nephropathy via inhibition of advanced glycation end products accumulation in STZ-nicotinamide induced diabetic rats

    doi: 10.1016/j.jtcme.2017.06.004

    Figure Lengend Snippet: HPLC chromatogram for DAE (A), DHE (B) standard Betulinic acid (C). Arrow indicates peak for betulinic acid.

    Article Snippet: 2.6 Quantification of betulinic acid by using HPLC Betulinic acid was quantified in DAE and DHE by high-performance liquid chromatography (HPLC) analysis using a Waters 2795 HPLC system with a UV detector.

    Techniques: High Performance Liquid Chromatography

    Sections of chromatograms (between retention times 2.5 and 6.5 min) of a cocktail of known metabolites ( a , mix1), untreated CE population cell lysate ( b , 3P), cell lysate from CE populations exposed to 250 ppm ( c , 3L2), 500 ppm ( d , 3L5), 1000 ppm ( e , 3L10) and 2000 ppm ( f , 3L20) lead acetate, obtained by CoulArray HPLC, using the CoulArray software. a Chromatogram of a cocktail of known metabolites (1 mg/ml each). Uric acid eluted at 3.60 min (min) with its primary channel being 5; 8-hydroxy guanine at 3.98 min, channels 5 and 6; xanthine (xan) at 4.28 min, channels 12 and 13; guanine at 4.87 min, channels 11 and 12; guanosine at 5.28 min, channels 14 and 15; xanthosine at 5.56 min, channels 14 and 15; and, tyrosine at 5.93 min, channels 11, 12 and 13. b Chromatogram of sample 3PA [untreated 14-day CE population (P), the third replicate of set A , normalization factor 0.26]. c Chromatogram of sample 3L2A [14-day 250 ppm lead acetate treated CE population (L2), the third replicate of set A , normalization factor 0.41]. d Chromatogram of sample 3L5A [14-day 500 ppm lead acetate treated CE population (L5), the third replicate of set A , normalization factor 0.54]. e Chromatogram of sample 3L10A [14-day 1000 ppm lead acetate treated CE population (L10), the third replicate of set A, normalization factor 0.63]. f Chromatogram of sample 3L20A [14-day 2000 ppm lead acetate treated CE population (L20), the third replicate of set A, normalization factor 0.68]. One can see differences when comparing the chromatograms from the untreated to the various levels of lead treated nematode populations

    Journal: Metabolomics

    Article Title: Metabolic profiling in Caenorhabditis elegans provides an unbiased approach to investigations of dosage dependent lead toxicity

    doi: 10.1007/s11306-012-0438-0

    Figure Lengend Snippet: Sections of chromatograms (between retention times 2.5 and 6.5 min) of a cocktail of known metabolites ( a , mix1), untreated CE population cell lysate ( b , 3P), cell lysate from CE populations exposed to 250 ppm ( c , 3L2), 500 ppm ( d , 3L5), 1000 ppm ( e , 3L10) and 2000 ppm ( f , 3L20) lead acetate, obtained by CoulArray HPLC, using the CoulArray software. a Chromatogram of a cocktail of known metabolites (1 mg/ml each). Uric acid eluted at 3.60 min (min) with its primary channel being 5; 8-hydroxy guanine at 3.98 min, channels 5 and 6; xanthine (xan) at 4.28 min, channels 12 and 13; guanine at 4.87 min, channels 11 and 12; guanosine at 5.28 min, channels 14 and 15; xanthosine at 5.56 min, channels 14 and 15; and, tyrosine at 5.93 min, channels 11, 12 and 13. b Chromatogram of sample 3PA [untreated 14-day CE population (P), the third replicate of set A , normalization factor 0.26]. c Chromatogram of sample 3L2A [14-day 250 ppm lead acetate treated CE population (L2), the third replicate of set A , normalization factor 0.41]. d Chromatogram of sample 3L5A [14-day 500 ppm lead acetate treated CE population (L5), the third replicate of set A , normalization factor 0.54]. e Chromatogram of sample 3L10A [14-day 1000 ppm lead acetate treated CE population (L10), the third replicate of set A, normalization factor 0.63]. f Chromatogram of sample 3L20A [14-day 2000 ppm lead acetate treated CE population (L20), the third replicate of set A, normalization factor 0.68]. One can see differences when comparing the chromatograms from the untreated to the various levels of lead treated nematode populations

    Article Snippet: CE cell lysates from each sample were subjected to HPLC analysis to generate chromatograms (CoulArray® software) and were assayed for total protein content using the Bradford protein assay procedure (BIO-RAD, P123236, Hercules, CA).

    Techniques: High Performance Liquid Chromatography, Software

    Evaluation of the intracellular shikonin concentration in shikonin-treated glioma cells. U87MG cells were treated with 2, 8, 50 and 100 μM of shikonin for 2 h. After treatment, cells were shattered by sonication and then centrifuged at 12,000 g for 10 min at 4°C. The cytosolic substances (supernatant) were evaluated the shikonin concentration detected by HPLC.

    Journal: PLoS ONE

    Article Title: An Oxidative Stress Mechanism of Shikonin in Human Glioma Cells

    doi: 10.1371/journal.pone.0094180

    Figure Lengend Snippet: Evaluation of the intracellular shikonin concentration in shikonin-treated glioma cells. U87MG cells were treated with 2, 8, 50 and 100 μM of shikonin for 2 h. After treatment, cells were shattered by sonication and then centrifuged at 12,000 g for 10 min at 4°C. The cytosolic substances (supernatant) were evaluated the shikonin concentration detected by HPLC.

    Article Snippet: HPLC quantitative analysis of shikonin in cells For HPLC analysis using with RP-C18 column (4.6 mm×250 mm, 5 μm, Merck, Germany), methanol–water (85∶15, v/v) was used as the mobile phase.

    Techniques: Concentration Assay, Sonication, High Performance Liquid Chromatography

    Effect of HMNQ and HQNO on the primary metabolome of E. coli . (A) Shown is the fold-change in the levels of 93 primary metabolites as a function of HMNQ or HQNO treatment, with respect to a no-treatment control, determined by HR-HPLC-MS. Three independent biological replicates for both drug-treated and untreated cells were used. The metabolites are sorted by fold-change. The color-coding is as follows: pyrimidine biosynthetic intermediates, red; NMPs, green; canonical amino acids, purple; NTPs, blue. (B) Biosynthetic pathway for pyrimidine nucleotides. HMNQ and HQNO inhibit DHODH, which results in accumulation of DHO and N-carbamoyl-L-aspartate (carbamoyl-Asp).

    Journal: Cell chemical biology

    Article Title: Synergy and Target Promiscuity Drive Structural Divergence in Bacterial Alkylquinolone Biosynthesis

    doi: 10.1016/j.chembiol.2017.08.024

    Figure Lengend Snippet: Effect of HMNQ and HQNO on the primary metabolome of E. coli . (A) Shown is the fold-change in the levels of 93 primary metabolites as a function of HMNQ or HQNO treatment, with respect to a no-treatment control, determined by HR-HPLC-MS. Three independent biological replicates for both drug-treated and untreated cells were used. The metabolites are sorted by fold-change. The color-coding is as follows: pyrimidine biosynthetic intermediates, red; NMPs, green; canonical amino acids, purple; NTPs, blue. (B) Biosynthetic pathway for pyrimidine nucleotides. HMNQ and HQNO inhibit DHODH, which results in accumulation of DHO and N-carbamoyl-L-aspartate (carbamoyl-Asp).

    Article Snippet: Peaks containing HMNQ, as judged by HPLC-MS analysis, were pooled, dried in vacuo and purified to homogeneity on an Agilent semi-preparative HPLC using an Eclipse XDB-C8 column (250 × 9.4mm, 5 μm) operating at 2.5 mL/min and an isocratic elution step of 60% MeCN (in H2 O).

    Techniques: High Performance Liquid Chromatography, Mass Spectrometry

    Nitration degrees for dissolved Bet v 1 nitrated by peroxynitrite (ONOO – ) in aqueous solution plotted against the molar ratio of nitrating agent and tyrosine residues (ONOO – /Y): (a) total nitration degrees (ND) determined by AAA, HPLC-DAD, and LC–MS/MS and (b) site-specific nitration degrees (ND Y ) determined by LC–MS/MS. Data points and error bars represent the arithmetic mean values and standard errors of eight experiments.

    Journal: Journal of Proteome Research

    Article Title: Nitration of the Birch Pollen Allergen Bet v 1.0101: Efficiency and Site-Selectivity of Liquid and Gaseous Nitrating Agents

    doi: 10.1021/pr401078h

    Figure Lengend Snippet: Nitration degrees for dissolved Bet v 1 nitrated by peroxynitrite (ONOO – ) in aqueous solution plotted against the molar ratio of nitrating agent and tyrosine residues (ONOO – /Y): (a) total nitration degrees (ND) determined by AAA, HPLC-DAD, and LC–MS/MS and (b) site-specific nitration degrees (ND Y ) determined by LC–MS/MS. Data points and error bars represent the arithmetic mean values and standard errors of eight experiments.

    Article Snippet: HPLC–DAD Analysis The protein solutions were analyzed using an HPLC–DAD system (Agilent Technologies 1200 series), as previously described.

    Techniques: Nitration, High Performance Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry

    Nitration degrees for Bet v 1 exposed to ozone and nitrogen dioxide (O 3 /NO 2 ) plotted against exposure time: (a) Total nitration degrees (ND) determined by HPLC–DAD for different types of samples and reaction conditions; data points and error bars represent the arithmetic mean values and standard errors of four experiments with filter samples and two experiments with liquid samples, respectively. The dashed lines and the gray shaded area show the results of kinetic model simulations, as indicated in the Figure and detailed in the Supporting Information (Section S4). The black dotted line represents the maximum ND for Bet v 1 based on the assumptions used in the model simulations. (b) Site-specific nitration degrees (ND Y ) determined by LC–MS/MS for filter samples exposed to 230 ppb of O 3 /NO 2 at a relative humidity of 92%.

    Journal: Journal of Proteome Research

    Article Title: Nitration of the Birch Pollen Allergen Bet v 1.0101: Efficiency and Site-Selectivity of Liquid and Gaseous Nitrating Agents

    doi: 10.1021/pr401078h

    Figure Lengend Snippet: Nitration degrees for Bet v 1 exposed to ozone and nitrogen dioxide (O 3 /NO 2 ) plotted against exposure time: (a) Total nitration degrees (ND) determined by HPLC–DAD for different types of samples and reaction conditions; data points and error bars represent the arithmetic mean values and standard errors of four experiments with filter samples and two experiments with liquid samples, respectively. The dashed lines and the gray shaded area show the results of kinetic model simulations, as indicated in the Figure and detailed in the Supporting Information (Section S4). The black dotted line represents the maximum ND for Bet v 1 based on the assumptions used in the model simulations. (b) Site-specific nitration degrees (ND Y ) determined by LC–MS/MS for filter samples exposed to 230 ppb of O 3 /NO 2 at a relative humidity of 92%.

    Article Snippet: HPLC–DAD Analysis The protein solutions were analyzed using an HPLC–DAD system (Agilent Technologies 1200 series), as previously described.

    Techniques: Nitration, High Performance Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry

    Nitration degrees for dissolved Bet v 1 nitrated by tetranitromethane (TNM) in solution plotted against the molar ratio of nitrating agent and tyrosine residues (TNM/Y): (a) total nitration degrees (ND) determined by AAA, HPLC–DAD, and LC–MS/MS and (b) site-specific nitration degrees (ND Y ) determined by LC–MS/MS. Data points and error bars represent the arithmetic mean values and standard errors of four chromatographic runs.

    Journal: Journal of Proteome Research

    Article Title: Nitration of the Birch Pollen Allergen Bet v 1.0101: Efficiency and Site-Selectivity of Liquid and Gaseous Nitrating Agents

    doi: 10.1021/pr401078h

    Figure Lengend Snippet: Nitration degrees for dissolved Bet v 1 nitrated by tetranitromethane (TNM) in solution plotted against the molar ratio of nitrating agent and tyrosine residues (TNM/Y): (a) total nitration degrees (ND) determined by AAA, HPLC–DAD, and LC–MS/MS and (b) site-specific nitration degrees (ND Y ) determined by LC–MS/MS. Data points and error bars represent the arithmetic mean values and standard errors of four chromatographic runs.

    Article Snippet: HPLC–DAD Analysis The protein solutions were analyzed using an HPLC–DAD system (Agilent Technologies 1200 series), as previously described.

    Techniques: Nitration, High Performance Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry

    Size exclusion and reversed-phase HPLC analysis of purified R0.6C. a Size exclusion chromatography was performed under native conditions in a phosphate buffer pH 6.7, to determine the amount of monomer in the sample. The peak corresponding to the monomer is indicated with the integrated area of the peak written above. b Reversed-phase HPLC–UV chromatograms recorded following analysis of purified protein batches. The peak at 17 min corresponds to monomeric R0.6C antigen

    Journal: Microbial Cell Factories

    Article Title: Construct design, production, and characterization of Plasmodium falciparum 48/45 R0.6C subunit protein produced in Lactococcus lactis as candidate vaccine

    doi: 10.1186/s12934-017-0710-0

    Figure Lengend Snippet: Size exclusion and reversed-phase HPLC analysis of purified R0.6C. a Size exclusion chromatography was performed under native conditions in a phosphate buffer pH 6.7, to determine the amount of monomer in the sample. The peak corresponding to the monomer is indicated with the integrated area of the peak written above. b Reversed-phase HPLC–UV chromatograms recorded following analysis of purified protein batches. The peak at 17 min corresponds to monomeric R0.6C antigen

    Article Snippet: SEC–HPLC and RP-HPLC analysis of R0.6C Native size exclusion high-performance liquid chromatography (SEC–HPLC) or reversed-phase HPLC (RP-HPLC) of intact R0.6C were performed using an Agilent 1100 Series HPLC System (Agilent Technologies, USA) equipped with a TSKgel G3000SWXL SEC column, 5 µm, 7.8 × 300 mm (Tosoh Bioscience, Japan) or equipped with a Vydac 214TP C4 reversed-phase column, 5 µm, 4.6 × 250 mm (The Separations Group, CA, US), respectively.

    Techniques: High Performance Liquid Chromatography, Purification, Size-exclusion Chromatography

    (a and b) Calibration plot of the khellin using different concentrations versus peak area for HPLC and HPTLC

    Journal: Journal of Pharmacy & Bioallied Sciences

    Article Title: Development and validation of high-performance liquid chromatography and high-performance thin-layer chromatography methods for the quantification of khellin in Ammi visnaga seed

    doi: 10.4103/0975-7406.168033

    Figure Lengend Snippet: (a and b) Calibration plot of the khellin using different concentrations versus peak area for HPLC and HPTLC

    Article Snippet: All other chemicals were of HPLC grade for HPLC analysis and analytical reagent grade for HPTLC analysis purchased from Merck Specialties Pvt.

    Techniques: High Performance Liquid Chromatography, High Performance Thin Layer Chromatography

    HPLC chromatograms of SAT I (A) and SAT IV (B) assays with calf spleen microsomes; calf spleen microsomes were extracted with Triton. The resulting extract was tested for SAT I (A) and SAT IV (B) activity by incubation with the respective BODIPY-labelled substrates for 1 h at 37°C. The presence of the respective activity was indicated by the presence of the product (peak 2) next to unconverted substrate (peak 1).

    Journal: PLoS ONE

    Article Title: A New Assay for Determining Ganglioside Sialyltransferase Activities Lactosylceramide-2,3-Sialyltransferase (SAT I) and Monosialylganglioside-2,3-Sialyltransferase (SAT IV)

    doi: 10.1371/journal.pone.0094206

    Figure Lengend Snippet: HPLC chromatograms of SAT I (A) and SAT IV (B) assays with calf spleen microsomes; calf spleen microsomes were extracted with Triton. The resulting extract was tested for SAT I (A) and SAT IV (B) activity by incubation with the respective BODIPY-labelled substrates for 1 h at 37°C. The presence of the respective activity was indicated by the presence of the product (peak 2) next to unconverted substrate (peak 1).

    Article Snippet: HPLC analysis of BODIPY gangliosides and unlabelled gangliosides The HPLC analysis was conducted on a Gilson system (Gilson, Middleton,Wisconsin, USA) with three pumps, a gradient mixer and an auto-sampler.

    Techniques: High Performance Liquid Chromatography, Activity Assay, Incubation

    HPLC chromatogram of the mixture resulting from the reaction with rat liver 2,3-sialyltransferase; BODIPY-GM1 (76 μM) and CMP-NANA (144 μM) were incubated with rat liver 2,3-sialyltransferase (1.4 mU) for one hour at 37°C. The sample was snap-frozen, freeze-dried and dissolved in chloroform/methanol. The solution was analysed by HPLC. The product (peak 2) eluted after the acceptor substrate (peak 1) with baseline separation.

    Journal: PLoS ONE

    Article Title: A New Assay for Determining Ganglioside Sialyltransferase Activities Lactosylceramide-2,3-Sialyltransferase (SAT I) and Monosialylganglioside-2,3-Sialyltransferase (SAT IV)

    doi: 10.1371/journal.pone.0094206

    Figure Lengend Snippet: HPLC chromatogram of the mixture resulting from the reaction with rat liver 2,3-sialyltransferase; BODIPY-GM1 (76 μM) and CMP-NANA (144 μM) were incubated with rat liver 2,3-sialyltransferase (1.4 mU) for one hour at 37°C. The sample was snap-frozen, freeze-dried and dissolved in chloroform/methanol. The solution was analysed by HPLC. The product (peak 2) eluted after the acceptor substrate (peak 1) with baseline separation.

    Article Snippet: HPLC analysis of BODIPY gangliosides and unlabelled gangliosides The HPLC analysis was conducted on a Gilson system (Gilson, Middleton,Wisconsin, USA) with three pumps, a gradient mixer and an auto-sampler.

    Techniques: High Performance Liquid Chromatography, Incubation

    HPLC chromatograms of BODIPY-GM3 (A) and BODIPY-GD1a (B) recovered from enzyme reactions; reaction products from the sialyltransferase reactions were purified by HPLC and analysed (top trace) showing mainly the product (peak 2) and only little amounts of starting material (peak 1). The samples were treated with sialidase overnight at 37°C (lower trace, approximately 10-fold dilution from treatment before) and analysed showing complete conversion to the starting material (peak 1).

    Journal: PLoS ONE

    Article Title: A New Assay for Determining Ganglioside Sialyltransferase Activities Lactosylceramide-2,3-Sialyltransferase (SAT I) and Monosialylganglioside-2,3-Sialyltransferase (SAT IV)

    doi: 10.1371/journal.pone.0094206

    Figure Lengend Snippet: HPLC chromatograms of BODIPY-GM3 (A) and BODIPY-GD1a (B) recovered from enzyme reactions; reaction products from the sialyltransferase reactions were purified by HPLC and analysed (top trace) showing mainly the product (peak 2) and only little amounts of starting material (peak 1). The samples were treated with sialidase overnight at 37°C (lower trace, approximately 10-fold dilution from treatment before) and analysed showing complete conversion to the starting material (peak 1).

    Article Snippet: HPLC analysis of BODIPY gangliosides and unlabelled gangliosides The HPLC analysis was conducted on a Gilson system (Gilson, Middleton,Wisconsin, USA) with three pumps, a gradient mixer and an auto-sampler.

    Techniques: High Performance Liquid Chromatography, Purification

    Role of PGF 2α in adrenal endocrine functions. A , ELISA quantification of PGF 2α release by chromaffin MPC862L cells untreated (control) or treated with 10 −6 M dexamethasone for 12 h (dex). B , HPLC quantification of dopamine secretion by MPC862L cells cultured in absence or presence of 10 −7 M cloprostenol (clo) (PGF 2α analogue) used either alone or in combination with 10 −6 M dexamethasone for 12 h (clo+dex). Values are the mean of 3 experiments ± S.D. *, P

    Journal: PLoS ONE

    Article Title: Aldo Keto Reductase 1B7 and Prostaglandin F2α Are Regulators of Adrenal Endocrine Functions

    doi: 10.1371/journal.pone.0007309

    Figure Lengend Snippet: Role of PGF 2α in adrenal endocrine functions. A , ELISA quantification of PGF 2α release by chromaffin MPC862L cells untreated (control) or treated with 10 −6 M dexamethasone for 12 h (dex). B , HPLC quantification of dopamine secretion by MPC862L cells cultured in absence or presence of 10 −7 M cloprostenol (clo) (PGF 2α analogue) used either alone or in combination with 10 −6 M dexamethasone for 12 h (clo+dex). Values are the mean of 3 experiments ± S.D. *, P

    Article Snippet: Analysis of catecholamine release Dopamine release was measured in 12 h culture media of MPC862L chromaffin line by HPLC analysis with the Chromsystems kit (München, Germany).

    Techniques: Enzyme-linked Immunosorbent Assay, High Performance Liquid Chromatography, Cell Culture