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Agilent technologies liquid chromatography tandem mass spectrometry
Wld S downregulates UCP2 expression and upregulates ATP levels through SIRT1. A : Wld S colocalized with SIRT1 in MIN6 cells. The stable cell lines expressing EGFP-Wld S were transiently transfected with pCMV-myc-SIRT1 and stained with anti-Myc antibody and DAPI. Scale bar, 5 μm. B : Wld S was coimmunoprecipitated with SIRT1 from the pancreatic lysates of Wld S mice. C : Wld S and its enzyme-dead mutant Wld S -H112A coimmunoprecipitated with SIRT1. The MIN6 cells stably expressing EGFP, EGFP-Wld S , or EGFP-Wld S -H112A were used for immunoprecipitation. UCP2 protein levels were also detected by Western blot. D and E : <t>Liquid</t> <t>chromatography-tandem</t> <t>mass</t> <t>spectrometry</t> analysis of NMN, NADP, NADPH, NAD, NADH, NA, and NAM extracted from pancreas of 9-week-old wild-type (WT) ( D ) or Wld S mice ( E ). *With significant difference. F : Quantification of the small molecules corresponding to D and E showed that NAD and NMN levels were upregulated in the pancreas of Wld S mice ( n = 4 for each genotype). G : Wld S repressed UCP2 promoter activity like SIRT1. UCP2 promoter activity was measured by luciferase assay in 293T cells transfected with pGL3-UCP2-Promoter and the indicated plasmids. H : Wld S downregulated UCP2 mRNA levels dependent on its enzyme activity. UCP2 mRNA level was measured by real-time PCR with MIN6 cell lines stably expressing EGFP, EGFP-Wld S , and EGFP-Wld S -H112A. I : Wld S downregulated UCP2 mRNA levels via SIRT1. UCP2 mRNA levels were determined by real-time PCR using islets isolated from mice with the indicated genotype ( n = 4 for each genotype). J : Wld S downregulated UCP2 protein levels via SIRT1. The protein levels in brown fat tissue with the indicated genotype were detected with SIRT1, Wld S , UCP2, and tubulin antibodies ( n = 3 for each genotype). K : Quantification of the UCP2 protein levels corresponding to J . L : Wld S increased ATP levels in primary cultured islets at the indicated glucose concentration ( n = 3). M : Wld S upregulated ATP level in islets via SIRT1. ATP levels were measured in islets with indicated genotypes at 2 mmol/L or 20 mmol/L glucose ( n = 3). * P
Liquid Chromatography Tandem Mass Spectrometry, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 93/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "WldS Enhances Insulin Transcription and Secretion via a SIRT1-Dependent Pathway and Improves Glucose Homeostasis"

Article Title: WldS Enhances Insulin Transcription and Secretion via a SIRT1-Dependent Pathway and Improves Glucose Homeostasis

Journal: Diabetes

doi: 10.2337/db11-0232

Wld S downregulates UCP2 expression and upregulates ATP levels through SIRT1. A : Wld S colocalized with SIRT1 in MIN6 cells. The stable cell lines expressing EGFP-Wld S were transiently transfected with pCMV-myc-SIRT1 and stained with anti-Myc antibody and DAPI. Scale bar, 5 μm. B : Wld S was coimmunoprecipitated with SIRT1 from the pancreatic lysates of Wld S mice. C : Wld S and its enzyme-dead mutant Wld S -H112A coimmunoprecipitated with SIRT1. The MIN6 cells stably expressing EGFP, EGFP-Wld S , or EGFP-Wld S -H112A were used for immunoprecipitation. UCP2 protein levels were also detected by Western blot. D and E : Liquid chromatography-tandem mass spectrometry analysis of NMN, NADP, NADPH, NAD, NADH, NA, and NAM extracted from pancreas of 9-week-old wild-type (WT) ( D ) or Wld S mice ( E ). *With significant difference. F : Quantification of the small molecules corresponding to D and E showed that NAD and NMN levels were upregulated in the pancreas of Wld S mice ( n = 4 for each genotype). G : Wld S repressed UCP2 promoter activity like SIRT1. UCP2 promoter activity was measured by luciferase assay in 293T cells transfected with pGL3-UCP2-Promoter and the indicated plasmids. H : Wld S downregulated UCP2 mRNA levels dependent on its enzyme activity. UCP2 mRNA level was measured by real-time PCR with MIN6 cell lines stably expressing EGFP, EGFP-Wld S , and EGFP-Wld S -H112A. I : Wld S downregulated UCP2 mRNA levels via SIRT1. UCP2 mRNA levels were determined by real-time PCR using islets isolated from mice with the indicated genotype ( n = 4 for each genotype). J : Wld S downregulated UCP2 protein levels via SIRT1. The protein levels in brown fat tissue with the indicated genotype were detected with SIRT1, Wld S , UCP2, and tubulin antibodies ( n = 3 for each genotype). K : Quantification of the UCP2 protein levels corresponding to J . L : Wld S increased ATP levels in primary cultured islets at the indicated glucose concentration ( n = 3). M : Wld S upregulated ATP level in islets via SIRT1. ATP levels were measured in islets with indicated genotypes at 2 mmol/L or 20 mmol/L glucose ( n = 3). * P
Figure Legend Snippet: Wld S downregulates UCP2 expression and upregulates ATP levels through SIRT1. A : Wld S colocalized with SIRT1 in MIN6 cells. The stable cell lines expressing EGFP-Wld S were transiently transfected with pCMV-myc-SIRT1 and stained with anti-Myc antibody and DAPI. Scale bar, 5 μm. B : Wld S was coimmunoprecipitated with SIRT1 from the pancreatic lysates of Wld S mice. C : Wld S and its enzyme-dead mutant Wld S -H112A coimmunoprecipitated with SIRT1. The MIN6 cells stably expressing EGFP, EGFP-Wld S , or EGFP-Wld S -H112A were used for immunoprecipitation. UCP2 protein levels were also detected by Western blot. D and E : Liquid chromatography-tandem mass spectrometry analysis of NMN, NADP, NADPH, NAD, NADH, NA, and NAM extracted from pancreas of 9-week-old wild-type (WT) ( D ) or Wld S mice ( E ). *With significant difference. F : Quantification of the small molecules corresponding to D and E showed that NAD and NMN levels were upregulated in the pancreas of Wld S mice ( n = 4 for each genotype). G : Wld S repressed UCP2 promoter activity like SIRT1. UCP2 promoter activity was measured by luciferase assay in 293T cells transfected with pGL3-UCP2-Promoter and the indicated plasmids. H : Wld S downregulated UCP2 mRNA levels dependent on its enzyme activity. UCP2 mRNA level was measured by real-time PCR with MIN6 cell lines stably expressing EGFP, EGFP-Wld S , and EGFP-Wld S -H112A. I : Wld S downregulated UCP2 mRNA levels via SIRT1. UCP2 mRNA levels were determined by real-time PCR using islets isolated from mice with the indicated genotype ( n = 4 for each genotype). J : Wld S downregulated UCP2 protein levels via SIRT1. The protein levels in brown fat tissue with the indicated genotype were detected with SIRT1, Wld S , UCP2, and tubulin antibodies ( n = 3 for each genotype). K : Quantification of the UCP2 protein levels corresponding to J . L : Wld S increased ATP levels in primary cultured islets at the indicated glucose concentration ( n = 3). M : Wld S upregulated ATP level in islets via SIRT1. ATP levels were measured in islets with indicated genotypes at 2 mmol/L or 20 mmol/L glucose ( n = 3). * P

Techniques Used: Expressing, Stable Transfection, Transfection, Staining, Mouse Assay, Mutagenesis, Immunoprecipitation, Western Blot, Liquid Chromatography, Mass Spectrometry, Activity Assay, Luciferase, Real-time Polymerase Chain Reaction, Isolation, Cell Culture, Concentration Assay

2) Product Images from "Analysis of 953 Human Proteins from a Mitochondrial HEK293 Fraction by Complexome Profiling"

Article Title: Analysis of 953 Human Proteins from a Mitochondrial HEK293 Fraction by Complexome Profiling

Journal: PLoS ONE

doi: 10.1371/journal.pone.0068340

Schematic overview of the complexome profiling approach. Protein complexes are separated according to size by blue native gel electrophoresis after which the gel lane is cut into gel slices at even distance. Each gel slice is separately processed by tryptic in-gel digestion and subsequently analyzed by liquid chromatography combined with online tandem mass spectrometry. In the final steps, the peptide identifications with according relative abundance from each individual LC-MS/MS analysis are combined to reconstruct the migration profile for each protein that span the complete length of the blue native separation. Please note that two subunits of the large red complex were also available as a smaller complex in this example to include proteins that form multiple complexes.
Figure Legend Snippet: Schematic overview of the complexome profiling approach. Protein complexes are separated according to size by blue native gel electrophoresis after which the gel lane is cut into gel slices at even distance. Each gel slice is separately processed by tryptic in-gel digestion and subsequently analyzed by liquid chromatography combined with online tandem mass spectrometry. In the final steps, the peptide identifications with according relative abundance from each individual LC-MS/MS analysis are combined to reconstruct the migration profile for each protein that span the complete length of the blue native separation. Please note that two subunits of the large red complex were also available as a smaller complex in this example to include proteins that form multiple complexes.

Techniques Used: Nucleic Acid Electrophoresis, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Migration

3) Product Images from "Pharmacokinetic and pharmacodynamic integration of enrofloxacin against Salmonella Enteritidis after administering to broiler chicken by per-oral and intravenous routes"

Article Title: Pharmacokinetic and pharmacodynamic integration of enrofloxacin against Salmonella Enteritidis after administering to broiler chicken by per-oral and intravenous routes

Journal: Journal of Veterinary Science

doi: 10.4142/jvs.2019.20.e15

Representative LC-MS/MS chromatograms of spiked sample (A) and standard (B) solutions. LC-MS/MS , liquid chromatography-tandem mass spectrometry.
Figure Legend Snippet: Representative LC-MS/MS chromatograms of spiked sample (A) and standard (B) solutions. LC-MS/MS , liquid chromatography-tandem mass spectrometry.

Techniques Used: Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Liquid Chromatography

4) Product Images from "Triplet therapy with afatinib, cetuximab, and bevacizumab induces deep remission in lung cancer cells harboring EGFR T790M in vivo"

Article Title: Triplet therapy with afatinib, cetuximab, and bevacizumab induces deep remission in lung cancer cells harboring EGFR T790M in vivo

Journal: Molecular Oncology

doi: 10.1002/1878-0261.12063

Mechanisms of the deep remission induced by triplet therapy. RPC ‐9 cells were used in in vitro and in vivo models. (A) Triplet therapy with gefitinib, cetuximab, and bevacizumab did not induce deep remission in xenograft tumors. (B) The concentration of afatinib in the xenograft tumors was assessed by liquid chromatography–tandem mass spectrometry ( LC ‐ MS / MS ). Cetuximab and bevacizumab did not increase the concentration of afatinib in xenograft tumors. Bars, SE ; n.s., not significant. (C–F) The xenograft tumors were treated for 1 week with the indicated drugs and collected for analysis. Afa, afatinib (10 mg·kg −1 , five times per week p.o.); Cet, cetuximab (0.1 mg per body, once a week i.p.); or Bev, bevacizumab (2 mg·kg −1 , twice a week i.p.). (C) The inhibitory effect on the EGFR signaling pathway in xenograft tumors was assessed by western blot. (D) The percent of CD 31‐positive cells in the xenograft tumors treated with indicated drugs. Bars, SE . * P
Figure Legend Snippet: Mechanisms of the deep remission induced by triplet therapy. RPC ‐9 cells were used in in vitro and in vivo models. (A) Triplet therapy with gefitinib, cetuximab, and bevacizumab did not induce deep remission in xenograft tumors. (B) The concentration of afatinib in the xenograft tumors was assessed by liquid chromatography–tandem mass spectrometry ( LC ‐ MS / MS ). Cetuximab and bevacizumab did not increase the concentration of afatinib in xenograft tumors. Bars, SE ; n.s., not significant. (C–F) The xenograft tumors were treated for 1 week with the indicated drugs and collected for analysis. Afa, afatinib (10 mg·kg −1 , five times per week p.o.); Cet, cetuximab (0.1 mg per body, once a week i.p.); or Bev, bevacizumab (2 mg·kg −1 , twice a week i.p.). (C) The inhibitory effect on the EGFR signaling pathway in xenograft tumors was assessed by western blot. (D) The percent of CD 31‐positive cells in the xenograft tumors treated with indicated drugs. Bars, SE . * P

Techniques Used: In Vitro, In Vivo, Concentration Assay, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Western Blot

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Article Snippet: .. The samples were then analyzed using a liquid chromatography-tandem mass spectrometry with an Agilent 1200 series high-performance liquid chromatography system and a 4000 Q Trap tandem mass spectrometer (Applied Biosystems). .. The data analysis was done with the Applied Biosystems Analyst software 1.4.2.

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Liquid Chromatography with Mass Spectroscopy:

Article Title: Dynamic Changes and Prognostic Value of Gut Microbiota-Dependent Trimethylamine-N-Oxide in Acute Ischemic Stroke
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Chromatography:

Article Title: WldS Enhances Insulin Transcription and Secretion via a SIRT1-Dependent Pathway and Improves Glucose Homeostasis
Article Snippet: .. The samples were then analyzed using a liquid chromatography-tandem mass spectrometry with an Agilent 1200 series high-performance liquid chromatography system and a 4000 Q Trap tandem mass spectrometer (Applied Biosystems). .. The data analysis was done with the Applied Biosystems Analyst software 1.4.2.

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Quantitation Assay:

Article Title: Loss of SLC25A11 causes suppression of NSCLC and melanoma tumor formation
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Isotope Dilution:

Article Title: Dynamic Changes and Prognostic Value of Gut Microbiota-Dependent Trimethylamine-N-Oxide in Acute Ischemic Stroke
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Mass Spectrometry:

Article Title: WldS Enhances Insulin Transcription and Secretion via a SIRT1-Dependent Pathway and Improves Glucose Homeostasis
Article Snippet: .. The samples were then analyzed using a liquid chromatography-tandem mass spectrometry with an Agilent 1200 series high-performance liquid chromatography system and a 4000 Q Trap tandem mass spectrometer (Applied Biosystems). .. The data analysis was done with the Applied Biosystems Analyst software 1.4.2.

Article Title: Triplet therapy with afatinib, cetuximab, and bevacizumab induces deep remission in lung cancer cells harboring EGFR T790M in vivo
Article Snippet: .. 2.6 Liquid chromatography–tandem mass spectrometry (LC‐MS/MS) conditions for afatinib quantification Chromatographic separation was performed using a high‐performance liquid chromatography system (Agilent 1100 series; Agilent, Santa Clara, CA, USA) and a CAPCELL PAK C18 MGIII S‐5 (100 mm × 2.0 mm; i.d., 3 μm; SHISEIDO, Tokyo, Japan) analytical column at 40 °C. .. The isocratic mobile phase consisted of mobile phase A (0.1% formic acid) and mobile phase B (methanol; 40 : 60, v/v) at a flow rate of 0.3 mL·min−1 .

Article Title: Loss of SLC25A11 causes suppression of NSCLC and melanoma tumor formation
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Article Title: Dynamic Changes and Prognostic Value of Gut Microbiota-Dependent Trimethylamine-N-Oxide in Acute Ischemic Stroke
Article Snippet: .. Plasma TMAO levels were quantified by stable isotope dilution liquid chromatography tandem mass spectrometry (6460 Series Triple Quadrupole LC/MS; Agilent) using a d9-(trimethyl)-labeled internal standard, as previously described ( ). .. In total, 50 μL of plasma was added to a 1.5 mL Eppendorf tube and mixed with 200 μL of 10 μmol/L internal standard composed of d9-TMAO in methanol.

Article Title: Pharmacokinetic and pharmacodynamic integration of enrofloxacin against Salmonella Enteritidis after administering to broiler chicken by per-oral and intravenous routes
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Article Title: Drosophila A virus is an unusual RNA virus with a T=3 icosahedral core and permuted RNA-dependent RNA polymerase
Article Snippet: .. Liquid chromatography–tandem mass spectrometry (LC-MS/MS) used an Agilent XCT-Ultra 6330 ion-trap mass spectrometer connected to an Agilent 1100 CapLC and ChipCube. .. Peptides were eluted and loaded onto the analytical capillary column (43 mm×75 μm inner diameter, also packed with 5 μm Zorbax 300SB-C18 particles) connected in line to the mass spectrometer with a flow of 600 nl min−1 .

Article Title: Influence of O Polysaccharides on Biofilm Development and Outer Membrane Vesicle Biogenesis in Pseudomonas aeruginosa PAO1
Article Snippet: .. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses were performed on an Agilent 1200 high-pressure liquid chromatography (HPLC) chromatograph interfaced with an Agilent UHD 6530 quadrupole time of flight (Q-TOF) mass spectrometer at the Mass Spectrometry Facility of the Advanced Analysis Centre, University of Guelph. .. A C18 column (Agilent Advance Bio Peptide map; 100 mm by 2.1 mm 2.7 μm) was used for chromatographic separation with solvents A (dH2 O with 0.1% formic acid) and B (acetonitrile with 0.1% formic acid).

Liquid Chromatography:

Article Title: Analysis of 953 Human Proteins from a Mitochondrial HEK293 Fraction by Complexome Profiling
Article Snippet: .. Liquid chromatography – tandem mass spectrometry Duplicate measurements for each gel slice were performed by nanoflow reversed-phase C18 liquid chromatography (Agilent 1100 series) coupled online to a 7T linear ion trap Fourier-Transform ion cyclotron resonance mass spectrometer (LTQ FT, Thermo Fisher Scientific) , . .. Chromatographic separations were performed using a 15 cm long×100 µm ID fused silica electrospray emitter (New Objective, PicoTip Emitter, FS360-100-8-N-5-C15) packed in-house at 100 bar with ReproSil-Pur C18AQ 3 µm 140 Å resin (Dr. Maisch) resuspended in methanol.

Article Title: Dynamic Changes and Prognostic Value of Gut Microbiota-Dependent Trimethylamine-N-Oxide in Acute Ischemic Stroke
Article Snippet: .. Plasma TMAO levels were quantified by stable isotope dilution liquid chromatography tandem mass spectrometry (6460 Series Triple Quadrupole LC/MS; Agilent) using a d9-(trimethyl)-labeled internal standard, as previously described ( ). .. In total, 50 μL of plasma was added to a 1.5 mL Eppendorf tube and mixed with 200 μL of 10 μmol/L internal standard composed of d9-TMAO in methanol.

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    Molecular design of Kα3-KFGGI via loop substitution. ( a ) Multiple sequence alignment of defensins (green bar), scorpion venom-derived K + channel toxins (red bar) and bifunctional scorpion toxins (pink bar), and the designed peptide (yellow bar). Secondary structure elements (α-helix, cylinder; β-strand, arrow) and disulfide bridges are extracted from the NMR structure of micasin (pdb entry 2LR5). Sequence sources: Mussel [ 23 ]; Insect [ 24 ]; Tick (GenPept No. ABW08118.1); Scorpion [ 25 ]; Micasin and Acasin [ 22 ]; Agitoxin-2 [ 26 ]; Kaliotoxin [ 27 ]; MeuTXKα3 and P30N [ 21 ]. The c-loop region is boxed, in which residues involved in antibacterial activity are colored according to their chemical properties (blue, basic; green, hydrophobic; cyan, polar) and an Asn essential for K + channel blocking activity is underlined once; ( b ) The computational structure of Kα3-KFGGI. Five residues introduced by loop substitution are indicated; ( c ) Surface potential distribution of Kα3-KFGGI, calculated by MOLMOL, with negative, positive, and neutral charge zones highlighted in red, blue, and white, respectively.

    Journal: Toxins

    Article Title: Loop Replacement Enhances the Ancestral Antibacterial Function of a Bifunctional Scorpion Toxin

    doi: 10.3390/toxins10060227

    Figure Lengend Snippet: Molecular design of Kα3-KFGGI via loop substitution. ( a ) Multiple sequence alignment of defensins (green bar), scorpion venom-derived K + channel toxins (red bar) and bifunctional scorpion toxins (pink bar), and the designed peptide (yellow bar). Secondary structure elements (α-helix, cylinder; β-strand, arrow) and disulfide bridges are extracted from the NMR structure of micasin (pdb entry 2LR5). Sequence sources: Mussel [ 23 ]; Insect [ 24 ]; Tick (GenPept No. ABW08118.1); Scorpion [ 25 ]; Micasin and Acasin [ 22 ]; Agitoxin-2 [ 26 ]; Kaliotoxin [ 27 ]; MeuTXKα3 and P30N [ 21 ]. The c-loop region is boxed, in which residues involved in antibacterial activity are colored according to their chemical properties (blue, basic; green, hydrophobic; cyan, polar) and an Asn essential for K + channel blocking activity is underlined once; ( b ) The computational structure of Kα3-KFGGI. Five residues introduced by loop substitution are indicated; ( c ) Surface potential distribution of Kα3-KFGGI, calculated by MOLMOL, with negative, positive, and neutral charge zones highlighted in red, blue, and white, respectively.

    Article Snippet: The released Kα3-KFGGI proteins were separated from GST by RP-HPLC on a C18 column (Zorbax 300SB-C18, 4.6 × 150 mm, 5 μm) (Agilent Technologies, Santa Clara, CA, USA).

    Techniques: Sequencing, Derivative Assay, Nuclear Magnetic Resonance, Activity Assay, Blocking Assay

    Characterization of recombinant Kα3-KFGGI. ( a ) RP-HPLC isolation of Kα3-KFGGI. The C 18 column was equilibrated with 0.05% TFA in water ( v / v ), and the EK-digested product was eluted from the column with a linear gradient from 0 to 60% acetonitrile in 0.05% TFA within 40 min; ( b ) MALDI-TOF MS determining the molecular mass of Kα3-KFGGI; ( c ) Circular dichroism spectra of Kα3-KFGGI. The spectra were recorded from 190 to 260 nm with a peptide concentration of 0.1 mg/mL in water. MeuTXKα3 and P30N were used as control [ 21 ].

    Journal: Toxins

    Article Title: Loop Replacement Enhances the Ancestral Antibacterial Function of a Bifunctional Scorpion Toxin

    doi: 10.3390/toxins10060227

    Figure Lengend Snippet: Characterization of recombinant Kα3-KFGGI. ( a ) RP-HPLC isolation of Kα3-KFGGI. The C 18 column was equilibrated with 0.05% TFA in water ( v / v ), and the EK-digested product was eluted from the column with a linear gradient from 0 to 60% acetonitrile in 0.05% TFA within 40 min; ( b ) MALDI-TOF MS determining the molecular mass of Kα3-KFGGI; ( c ) Circular dichroism spectra of Kα3-KFGGI. The spectra were recorded from 190 to 260 nm with a peptide concentration of 0.1 mg/mL in water. MeuTXKα3 and P30N were used as control [ 21 ].

    Article Snippet: The released Kα3-KFGGI proteins were separated from GST by RP-HPLC on a C18 column (Zorbax 300SB-C18, 4.6 × 150 mm, 5 μm) (Agilent Technologies, Santa Clara, CA, USA).

    Techniques: Recombinant, High Performance Liquid Chromatography, Isolation, Mass Spectrometry, Concentration Assay

    Mass spectra of intact form of GFPuv form I and form II. Mass spectra obtained by ESI-MS measurement for GFPuv form I ( top ) and form II ( middle ) were shown. The theoretical mass spectrum of GFPuv after the formation of the enol form of chromophore is shown ( bottom ).

    Journal: Scientific Reports

    Article Title: Specific modification at the C-terminal lysine residue of the green fluorescent protein variant, GFPuv, expressed in Escherichia coli

    doi: 10.1038/s41598-019-41309-8

    Figure Lengend Snippet: Mass spectra of intact form of GFPuv form I and form II. Mass spectra obtained by ESI-MS measurement for GFPuv form I ( top ) and form II ( middle ) were shown. The theoretical mass spectrum of GFPuv after the formation of the enol form of chromophore is shown ( bottom ).

    Article Snippet: Mass Spectrometric Analysis of the Recombinant GFPuv An electrospray ionization-mass spectrometry (ESI-MS) analysis of intact GFPuv forms I and II was carried out, using an HPLC-Chip/QTOF (G6520 and G4240, Agilent Technologies, Santa Clara, CA, USA).

    Techniques: Mass Spectrometry

    Determination of the modification type and site in GFPuv form II. ( A ) Liquid chromatography profiles of trypsin digests of GFPuv form I ( top ) and form II ( bottom ). A pair of peaks showing the retention time difference in GFPuv form I and form II digests is indicated by arrows. ( B ) ESI-MS spectrum of the H 231 GMDELYK 238 fragment ions from GFPuv form II. ( C ) Comparison of the observed mass of the H 231 GMDELYK 238 fragment ions from GFPuv form II with the theoretical mass spectra for the fragment containing the candidate modifications. ( D ) Schematic illustration of GFPuv form I and form II.

    Journal: Scientific Reports

    Article Title: Specific modification at the C-terminal lysine residue of the green fluorescent protein variant, GFPuv, expressed in Escherichia coli

    doi: 10.1038/s41598-019-41309-8

    Figure Lengend Snippet: Determination of the modification type and site in GFPuv form II. ( A ) Liquid chromatography profiles of trypsin digests of GFPuv form I ( top ) and form II ( bottom ). A pair of peaks showing the retention time difference in GFPuv form I and form II digests is indicated by arrows. ( B ) ESI-MS spectrum of the H 231 GMDELYK 238 fragment ions from GFPuv form II. ( C ) Comparison of the observed mass of the H 231 GMDELYK 238 fragment ions from GFPuv form II with the theoretical mass spectra for the fragment containing the candidate modifications. ( D ) Schematic illustration of GFPuv form I and form II.

    Article Snippet: Mass Spectrometric Analysis of the Recombinant GFPuv An electrospray ionization-mass spectrometry (ESI-MS) analysis of intact GFPuv forms I and II was carried out, using an HPLC-Chip/QTOF (G6520 and G4240, Agilent Technologies, Santa Clara, CA, USA).

    Techniques: Modification, Liquid Chromatography, Mass Spectrometry

    Schemes of flavan-3-ols (monomers) and dimeric proanthocyanidins: ( a ) four types of stereo configurations of different monomeric flavan-3-ols, such as (+)-catechin, (−)-epicatechin, (−)-epigallocatechin, and (−)-epicatechin; ( b ) two examples of dimeric proanthocyanidins, procyanidin A2 (A-type) and procyanidin B1 (B-type).

    Journal: Metabolites

    Article Title: HPLC-qTOF-MS/MS-Based Profiling of Flavan-3-ols and Dimeric Proanthocyanidins in Berries of Two Muscadine Grape Hybrids FLH 13-11 and FLH 17-66

    doi: 10.3390/metabo8040057

    Figure Lengend Snippet: Schemes of flavan-3-ols (monomers) and dimeric proanthocyanidins: ( a ) four types of stereo configurations of different monomeric flavan-3-ols, such as (+)-catechin, (−)-epicatechin, (−)-epigallocatechin, and (−)-epicatechin; ( b ) two examples of dimeric proanthocyanidins, procyanidin A2 (A-type) and procyanidin B1 (B-type).

    Article Snippet: The mobile phase solvents were composed of 1% acetic acid in water (solvent A: 1% HPLC grade acetic acid in LC-MS grade water) and 100% acetonitrile (solvent B) (LC-MS grade), which formed a gradient solvent system to separate flavan-3-ols and oligomeric PAs in an Elipes XDB-C18 analytical column (250 × 4.6 mm, 5 µM, 25 °C, Agilent).

    Techniques:

    Comparison of metabolite peak profiles between berry extracts of FLH 13-11 and FLH 17-66 in two years and nine standards. Metabolite peaks detected by HPLC were recorded at 280 nm. ( a ) Two chromatograms show peaks annotated to be flavan-3-ols and dimeric PAs in berries of FLH 13-11 from the 2011 and 2012 cropping years. ( b ) Two chromatograms show peaks annotated to be flavan-3-ols and dimeric PAs in berries of FLH 17-66 from the 2011 and 2012 cropping years. ( c ) A chromatogram shows retention times of nine standards including five flavan-3-ol aglycones, two conjugates, and two dimeric PA standards (procyanidin B1 and B2). Compound peaks detected in one variety only and two varieties are highlighted with red and black color, respectively. Standards and abbreviations, (+)-catechin (Cat), (−)-epicatechin (EpiCat), (−)-gallocatechin (gCat), (−)-epigallocatechin (EpigCat), (−)-catechin gallate (CatG), (−)-epigallocatechin gallate (EpigCatG), (−)-gallocatechin gallate (gCatG), procyanidin B1 (Proc B1), and procyanidin B2 (Proc B2). Abbreviation for compounds annotated by HPLC-qTOF-MS/MS analysis described below, (+)-Cat3Glu: (+)-catechin 3-glucoside, (−)-Cat3Glu: (−)-catechin 3-glucoside, (+)-EpiCat: (+)-epicatechin, (+)-gCat3Glu: (+)-gallocatechin 3-glucoside, (−)-gCat3Glu: (−)-gallocatechin 3-glucoside, (+)-EpigCat3Glu: (+)-epigallocatechin 3-glucuside, (−)-EpigCat3Glu: (−)-epigallocatechin 3-glucoside, OMe(+)-CatG: Methyl- O -(+)-catechin-3-gallate, OMe(−)-CatG: Methyl- O -(−)-catechin-3-gallate, OMe(−)-gCatG: Methyl- O -(−)-gallocatechin-3-gallate, OMe(+)-EpigCatG: Methyl- O -(−)-epigallocatechin-3-gallate, OMe(−)-EpigCatG: Methyl- O -(−)-epigallocatechin-3-gallate, and OMe(+/−)-EpiCatG: Methyl- O -(+/−)-epicatechin-3-gallate. Proc B4, B5, B6, B7, B8, and A2: procyanidin B4, B5, B6, B7, B8, and A2.

    Journal: Metabolites

    Article Title: HPLC-qTOF-MS/MS-Based Profiling of Flavan-3-ols and Dimeric Proanthocyanidins in Berries of Two Muscadine Grape Hybrids FLH 13-11 and FLH 17-66

    doi: 10.3390/metabo8040057

    Figure Lengend Snippet: Comparison of metabolite peak profiles between berry extracts of FLH 13-11 and FLH 17-66 in two years and nine standards. Metabolite peaks detected by HPLC were recorded at 280 nm. ( a ) Two chromatograms show peaks annotated to be flavan-3-ols and dimeric PAs in berries of FLH 13-11 from the 2011 and 2012 cropping years. ( b ) Two chromatograms show peaks annotated to be flavan-3-ols and dimeric PAs in berries of FLH 17-66 from the 2011 and 2012 cropping years. ( c ) A chromatogram shows retention times of nine standards including five flavan-3-ol aglycones, two conjugates, and two dimeric PA standards (procyanidin B1 and B2). Compound peaks detected in one variety only and two varieties are highlighted with red and black color, respectively. Standards and abbreviations, (+)-catechin (Cat), (−)-epicatechin (EpiCat), (−)-gallocatechin (gCat), (−)-epigallocatechin (EpigCat), (−)-catechin gallate (CatG), (−)-epigallocatechin gallate (EpigCatG), (−)-gallocatechin gallate (gCatG), procyanidin B1 (Proc B1), and procyanidin B2 (Proc B2). Abbreviation for compounds annotated by HPLC-qTOF-MS/MS analysis described below, (+)-Cat3Glu: (+)-catechin 3-glucoside, (−)-Cat3Glu: (−)-catechin 3-glucoside, (+)-EpiCat: (+)-epicatechin, (+)-gCat3Glu: (+)-gallocatechin 3-glucoside, (−)-gCat3Glu: (−)-gallocatechin 3-glucoside, (+)-EpigCat3Glu: (+)-epigallocatechin 3-glucuside, (−)-EpigCat3Glu: (−)-epigallocatechin 3-glucoside, OMe(+)-CatG: Methyl- O -(+)-catechin-3-gallate, OMe(−)-CatG: Methyl- O -(−)-catechin-3-gallate, OMe(−)-gCatG: Methyl- O -(−)-gallocatechin-3-gallate, OMe(+)-EpigCatG: Methyl- O -(−)-epigallocatechin-3-gallate, OMe(−)-EpigCatG: Methyl- O -(−)-epigallocatechin-3-gallate, and OMe(+/−)-EpiCatG: Methyl- O -(+/−)-epicatechin-3-gallate. Proc B4, B5, B6, B7, B8, and A2: procyanidin B4, B5, B6, B7, B8, and A2.

    Article Snippet: The mobile phase solvents were composed of 1% acetic acid in water (solvent A: 1% HPLC grade acetic acid in LC-MS grade water) and 100% acetonitrile (solvent B) (LC-MS grade), which formed a gradient solvent system to separate flavan-3-ols and oligomeric PAs in an Elipes XDB-C18 analytical column (250 × 4.6 mm, 5 µM, 25 °C, Agilent).

    Techniques: High Performance Liquid Chromatography, Mass Spectrometry

    LC-UV-MS-MS-based lipidomics: authentic and synthetic eicosanoids and docosanoids. LM lipidomics was conducted with an ABI Qtrap using MRM IDA EPI method (See text for details). A) Total ion chromatogram of 12 authentic and synthetic LM; Chromatogram of precursor/product ion pairs m/z 349 > 195 for RvE1 and m/z 375 > 141 for RvD1. B) Demonstration of online UV spectra for RvE1 and RvD1. MS-MS spectra of C) RvD1, and D) RvE1.

    Journal: Current Protocols in Immunology

    Article Title: Metabolomics-Lipidomics of Eicosanoids and Docosanoids Generated By Phagocytes

    doi: 10.1002/0471142735.im1426s95

    Figure Lengend Snippet: LC-UV-MS-MS-based lipidomics: authentic and synthetic eicosanoids and docosanoids. LM lipidomics was conducted with an ABI Qtrap using MRM IDA EPI method (See text for details). A) Total ion chromatogram of 12 authentic and synthetic LM; Chromatogram of precursor/product ion pairs m/z 349 > 195 for RvE1 and m/z 375 > 141 for RvD1. B) Demonstration of online UV spectra for RvE1 and RvD1. MS-MS spectra of C) RvD1, and D) RvE1.

    Article Snippet: Typical lipidomic/metabolomic analysis for eicosanoids and docosanoids can use an Agilent 1100 series HPLC coupled with an ABI 3200 Qtrap mass spectrometer equipped with an Agilent Eclipse Plus C18 column (4.6mm × 50mm × 1.8μm).

    Techniques: Mass Spectrometry