lc ms analysis  (Thermo Fisher)


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    Structured Review

    Thermo Fisher lc ms analysis
    Lc Ms Analysis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lc ms analysis/product/Thermo Fisher
    Average 98 stars, based on 28 article reviews
    Price from $9.99 to $1999.99
    lc ms analysis - by Bioz Stars, 2020-02
    98/100 stars

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    Related Articles

    High Performance Liquid Chromatography:

    Article Title: Biochemical assessment of red blood cells during storage by 1H nuclear magnetic resonance spectroscopy. Identification of a biomarker of their level of protection against oxidative stress
    Article Snippet: .. Liquid chromatography mass spectrometry analysis of the NMR samples of RBC supernant and lysates was performed using a LTQ Orbitrap XL (Thermo Scientific, Waltham, MA, USA) coupled to a HPLC Dionex Ultimate 3000. .. The liquid chromatography separation was carried out on Aeris peptide 3.6u XB-C18 column (150×2.1 mm) (Phenomenex, Torrance, CA, USA).

    Article Title: The corrinoid cofactor of reductive dehalogenases affects dechlorination rates and extents in organohalide-respiring Dehalococcoides mccartyi
    Article Snippet: Paragraph title: HPLC and liquid chromatography–mass spectrometry analysis ... Liquid chromatography–mass spectrometry analysis was performed using a Dionex Ultimate 3000 system (Thermo Fisher Scientific) with an inline DAD fitted to an Exactive Plus Orbitrap Mass Spectrometer with an electrospray ionization source (Thermo Fisher Scientific).

    Article Title: mTORC1 Activation Requires DRAM-1 by Facilitating Lysosomal Amino Acid Efflux
    Article Snippet: .. LC-MS Analysis A Q Exactive Orbitrap mass spectrometer (Thermo Scientific, Waltham, MA, USA) was used together with a Thermo Ultimate 3000 HPLC system. .. The HPLC setup consisted of a ZIC-pHILIC column (SeQuant, 150 × 2.1mm, 5 μm, Merck KGaA, Darmstadt, Germany), with a ZIC-pHILIC guard column (SeQuant, 20 × 2.1mm) and an initial mobile phase of 20% 20mM ammonium carbonate, pH 9.4, and 80% acetonitrile.

    Article Title: A proteomic chronology of gene expression through the cell cycle in human myeloid leukemia cells
    Article Snippet: .. Liquid chromatography mass spectrometry analysis (LC-MS/MS) For tryptic digests, including tryptic + Lys-C double digests, peptide chromatography was performed using a Dionex RSLCnano HPLC. ..

    Flow Cytometry:

    Article Title: Extraction and biomolecular analysis of dermal interstitial fluid collected with hollow microneedles
    Article Snippet: Liquid chromatography mass spectrometry analysis (LC-MS/MS) Each pH 10 fraction was re-suspended in 20 µl of acetonitrile/water/formic acid 5/95/0.5 (v/v/v) and 2 µl was analyzed in a nanoLC system (Dionex, Ultimate 3000) coupled to a Q Exactive plus mass spectrometer (Thermo Fisher Scientific, Waltham, MA). .. Peptides were loaded on a PepMap trap 300 um x 5 mm C18, 100 A (Thermo Fisher Scientific, Waltham, MA) at flow rate of 10 µl per min for 5 min. Peptides were then separated on an EASYspray C18 75 µm x 50 cm analytical column over a 182-min run at flow rate of 300 nl/min using mobile phase A of 0.1% formic acid in water and mobile phase B of acetonitrile/water/formic acid 80/20/0.1 (v/v/v).

    Article Title: ProTargetMiner as a proteome signature library of anticancer molecules for functional discovery
    Article Snippet: .. LC-MS analysis Samples were loaded with buffer A (0.1% FA in water) onto a 50 cm EASY-Spray column (75 µm internal diameter, packed with PepMap C18, 2 µm beads, 100 Å pore size; Cat#ES803) connected to the EASY-nLC 1000 (Thermo; Cat#LC120) and eluted with a buffer B (98% ACN, 0.1% FA, 2% H2 O) gradient from 2 to 35% of at a flow rate of 250 nL min−1 . .. Mass spectra were acquired with an Orbitrap Q Exactive Plus mass spectrometer (Thermo; Cat# IQLAAEGAAPFALGMBDK) in the data-dependent mode with MS1 scan at 70,000 resolution, and MS2 at 35,000, in the m/z range from 375 to 1400.

    Article Title: The corrinoid cofactor of reductive dehalogenases affects dechlorination rates and extents in organohalide-respiring Dehalococcoides mccartyi
    Article Snippet: For HPLC analysis, 20 μl samples were injected onto an Eclipse XDB-C18 column (5 μm, 4.6 × 150 mm) (Agilent) and separated with a flow rate of 1 ml per min at 30 °C using 0.1% (v/v) formic acid (⩾88%, w/v) in water (eluent A) and 0.1% (v/v) formic acid in methanol (eluent B) as mobile phases. .. Liquid chromatography–mass spectrometry analysis was performed using a Dionex Ultimate 3000 system (Thermo Fisher Scientific) with an inline DAD fitted to an Exactive Plus Orbitrap Mass Spectrometer with an electrospray ionization source (Thermo Fisher Scientific).

    Article Title: mTORC1 Activation Requires DRAM-1 by Facilitating Lysosomal Amino Acid Efflux
    Article Snippet: LC-MS Analysis A Q Exactive Orbitrap mass spectrometer (Thermo Scientific, Waltham, MA, USA) was used together with a Thermo Ultimate 3000 HPLC system. .. Cell and media extracts (5 μl) were injected and metabolites were separated over a 15 minute mobile phase gradient, decreasing the acetonitrile content to 20%, at a flow rate of 200μL/min and a column temperature of 45°C.

    Article Title: UGT85A84 Catalyzes the Glycosylation of Aromatic Monoterpenes in Osmanthus fragrans Lour. Flowers
    Article Snippet: LC-MS analysis was performed in the Thermo Scientific™ UltiMate™ 3000RS system. .. Chromatographic separation was achieved by gradient elution at a flow rate of 0.3 ml/min.

    Article Title: A proteomic chronology of gene expression through the cell cycle in human myeloid leukemia cells
    Article Snippet: Liquid chromatography mass spectrometry analysis (LC-MS/MS) For tryptic digests, including tryptic + Lys-C double digests, peptide chromatography was performed using a Dionex RSLCnano HPLC. .. The linear gradient began with 5% B to 35% B over 156 min with a constant flow of 300 nl/min.

    Article Title: Pharmacokinetic Study of Bioactive Glycopeptide from Strongylocentrotus droebachiensis After Intranasal Administration to Rats Using Biomarker Approach
    Article Snippet: The flow-through was collected as it contains peptides with a size of up to around 10 kDa. .. Before LC-MS analysis, peptide extracts were desalted and cleaned up using C18 spin tips (Pierce, Waltham, MS, USA) according to the manufacturer’s protocol.

    Incubation:

    Article Title: The function of the PduJ microcompartment shell protein is determined by the genomic position of its encoding gene
    Article Snippet: Trypsin (proteomics grade, Promega) was added to the gel pieces and incubated for 16–18 h according to a published procedure ( ). .. The trypsin digested peptides were extracted and subjected to liquid chromatography mass spectrometry analysis according to the manufacturer's instructions (Thermo, Q Exactive™ Hybrid Quadrupole-Obrbitrap Mass Spectrometer).

    Sample Prep:

    Article Title: Loss of NAD-Dependent Protein Deacetylase Sirtuin-2 Alters Mitochondrial Protein Acetylation and Dysregulates Mitophagy
    Article Snippet: Paragraph title: Mass spectrometry sample preparation and analysis ... Liquid chromatography-mass spectrometry analysis of the peptides was performed using an LTQ-Orbitrap mass spectrometer (Thermo Scientific) that was equipped with a nanospray source and an Eksigent NanoLC 1D Plus and AS1 Autosampler.

    Mass Spectrometry:

    Article Title: Extraction and biomolecular analysis of dermal interstitial fluid collected with hollow microneedles
    Article Snippet: .. Liquid chromatography mass spectrometry analysis (LC-MS/MS) Each pH 10 fraction was re-suspended in 20 µl of acetonitrile/water/formic acid 5/95/0.5 (v/v/v) and 2 µl was analyzed in a nanoLC system (Dionex, Ultimate 3000) coupled to a Q Exactive plus mass spectrometer (Thermo Fisher Scientific, Waltham, MA). .. Peptides were loaded on a PepMap trap 300 um x 5 mm C18, 100 A (Thermo Fisher Scientific, Waltham, MA) at flow rate of 10 µl per min for 5 min. Peptides were then separated on an EASYspray C18 75 µm x 50 cm analytical column over a 182-min run at flow rate of 300 nl/min using mobile phase A of 0.1% formic acid in water and mobile phase B of acetonitrile/water/formic acid 80/20/0.1 (v/v/v).

    Article Title: HLA-F Allele-Specific Peptide Restriction Represents an Exceptional Proteomic Footprint
    Article Snippet: .. The LC/MS analysis was performed using a Dionex Ultimate 3000 high-performance LC system and a LTQ Orbitrap Lumos Mass Spectrometer (Thermo Fisher Scientific) in data-dependent acquisition (DDA) mode. ..

    Article Title: Profiling the effect of nafcillin on HA-MRSA D712 using bacteriological and physiological media
    Article Snippet: Paragraph title: Untargeted liquid chromatography mass spectrometry data acquisition ... For LC/MS analysis, samples were subjected to chromatographic separation using an UltiMate 3000 UHPLC system (Thermo Scientific).

    Article Title: ProTargetMiner as a proteome signature library of anticancer molecules for functional discovery
    Article Snippet: LC-MS analysis Samples were loaded with buffer A (0.1% FA in water) onto a 50 cm EASY-Spray column (75 µm internal diameter, packed with PepMap C18, 2 µm beads, 100 Å pore size; Cat#ES803) connected to the EASY-nLC 1000 (Thermo; Cat#LC120) and eluted with a buffer B (98% ACN, 0.1% FA, 2% H2 O) gradient from 2 to 35% of at a flow rate of 250 nL min−1 . .. Mass spectra were acquired with an Orbitrap Q Exactive Plus mass spectrometer (Thermo; Cat# IQLAAEGAAPFALGMBDK) in the data-dependent mode with MS1 scan at 70,000 resolution, and MS2 at 35,000, in the m/z range from 375 to 1400.

    Article Title: The function of the PduJ microcompartment shell protein is determined by the genomic position of its encoding gene
    Article Snippet: .. The trypsin digested peptides were extracted and subjected to liquid chromatography mass spectrometry analysis according to the manufacturer's instructions (Thermo, Q Exactive™ Hybrid Quadrupole-Obrbitrap Mass Spectrometer). .. Tandem mass spectra were searched against the known protein sequence from S. enterica serovar Typhimurium LT2 using both MASCOT and Sequest HT (Thermo Proteome discoverer software from Thermo Fisher) with maximum mixed cleavage sites 2, fixed carbamidomethylation and variable methionine oxidation.

    Article Title: Biochemical assessment of red blood cells during storage by 1H nuclear magnetic resonance spectroscopy. Identification of a biomarker of their level of protection against oxidative stress
    Article Snippet: .. Liquid chromatography mass spectrometry analysis of the NMR samples of RBC supernant and lysates was performed using a LTQ Orbitrap XL (Thermo Scientific, Waltham, MA, USA) coupled to a HPLC Dionex Ultimate 3000. .. The liquid chromatography separation was carried out on Aeris peptide 3.6u XB-C18 column (150×2.1 mm) (Phenomenex, Torrance, CA, USA).

    Article Title: The corrinoid cofactor of reductive dehalogenases affects dechlorination rates and extents in organohalide-respiring Dehalococcoides mccartyi
    Article Snippet: .. Liquid chromatography–mass spectrometry analysis was performed using a Dionex Ultimate 3000 system (Thermo Fisher Scientific) with an inline DAD fitted to an Exactive Plus Orbitrap Mass Spectrometer with an electrospray ionization source (Thermo Fisher Scientific). .. For liquid chromatography–DAD–mass spectrometry analysis, 10 μl aliquots of each sample were injected onto a Kinetex XB-C18 column (2.6 μm, 2.1 × 100 mm) (Phenomenex, Torrance, CA, USA) and separated at a flow rate of 0.2 ml per min at 30 °C using 0.1% formic acid in water (eluent A) and 0.1% formic acid in acetonitrile (eluent B) as mobile phases.

    Article Title: A Resveratrol Analogue Promotes ERKMAPK
    Article Snippet: .. Liquid chromatography–mass spectrometry analysis was carried out on a Q-Exactive mass spectrometer coupled to an HTC autosampler and Accela pump (all from Thermo Fisher Scientific, Waltham, MA). .. Twenty-five microliters of the standard or extracts were injected onto an Ascentis Express C18 column (15 × 3 mm; 2.7 µ m; Supelco, Sigma-Aldrich).

    Article Title: mTORC1 Activation Requires DRAM-1 by Facilitating Lysosomal Amino Acid Efflux
    Article Snippet: .. LC-MS Analysis A Q Exactive Orbitrap mass spectrometer (Thermo Scientific, Waltham, MA, USA) was used together with a Thermo Ultimate 3000 HPLC system. .. The HPLC setup consisted of a ZIC-pHILIC column (SeQuant, 150 × 2.1mm, 5 μm, Merck KGaA, Darmstadt, Germany), with a ZIC-pHILIC guard column (SeQuant, 20 × 2.1mm) and an initial mobile phase of 20% 20mM ammonium carbonate, pH 9.4, and 80% acetonitrile.

    Article Title: A proteomic chronology of gene expression through the cell cycle in human myeloid leukemia cells
    Article Snippet: .. Liquid chromatography mass spectrometry analysis (LC-MS/MS) For tryptic digests, including tryptic + Lys-C double digests, peptide chromatography was performed using a Dionex RSLCnano HPLC. ..

    Article Title: Pharmacokinetic Study of Bioactive Glycopeptide from Strongylocentrotus droebachiensis After Intranasal Administration to Rats Using Biomarker Approach
    Article Snippet: .. Before LC-MS analysis, peptide extracts were desalted and cleaned up using C18 spin tips (Pierce, Waltham, MS, USA) according to the manufacturer’s protocol. .. The flow-through peptide sample was dissolved in 15 µL 0.1% trifluoroacetic acid (TFA, Fisher Scientific, Waltham, MS, USA) and 6 µL was injected into the LC-MS system.

    Article Title: Loss of NAD-Dependent Protein Deacetylase Sirtuin-2 Alters Mitochondrial Protein Acetylation and Dysregulates Mitophagy
    Article Snippet: .. Liquid chromatography-mass spectrometry analysis of the peptides was performed using an LTQ-Orbitrap mass spectrometer (Thermo Scientific) that was equipped with a nanospray source and an Eksigent NanoLC 1D Plus and AS1 Autosampler. ..

    Article Title: The integration of metabolome and proteome reveals bioactive polyphenols and hispidin in ARTP mutagenized Phellinus baumii
    Article Snippet: .. LC/MS analysis and statistical analysis Refering to previous research method , the ACQUITY UHPLC system Ultimate 3000 (Thermo Fisher Scientific, Waltham, MA, USA) coupled with LTQ Orbitrap MS (Thermo Fisher Scientific, Waltham, MA, USA) was used for metabolic profiling in both ESI+ and ESI− ion modes. ..

    Chromatography:

    Article Title: The corrinoid cofactor of reductive dehalogenases affects dechlorination rates and extents in organohalide-respiring Dehalococcoides mccartyi
    Article Snippet: .. Liquid chromatography–mass spectrometry analysis was performed using a Dionex Ultimate 3000 system (Thermo Fisher Scientific) with an inline DAD fitted to an Exactive Plus Orbitrap Mass Spectrometer with an electrospray ionization source (Thermo Fisher Scientific). .. For liquid chromatography–DAD–mass spectrometry analysis, 10 μl aliquots of each sample were injected onto a Kinetex XB-C18 column (2.6 μm, 2.1 × 100 mm) (Phenomenex, Torrance, CA, USA) and separated at a flow rate of 0.2 ml per min at 30 °C using 0.1% formic acid in water (eluent A) and 0.1% formic acid in acetonitrile (eluent B) as mobile phases.

    Article Title: A Resveratrol Analogue Promotes ERKMAPK
    Article Snippet: .. Liquid chromatography–mass spectrometry analysis was carried out on a Q-Exactive mass spectrometer coupled to an HTC autosampler and Accela pump (all from Thermo Fisher Scientific, Waltham, MA). .. Twenty-five microliters of the standard or extracts were injected onto an Ascentis Express C18 column (15 × 3 mm; 2.7 µ m; Supelco, Sigma-Aldrich).

    Article Title: A proteomic chronology of gene expression through the cell cycle in human myeloid leukemia cells
    Article Snippet: .. Liquid chromatography mass spectrometry analysis (LC-MS/MS) For tryptic digests, including tryptic + Lys-C double digests, peptide chromatography was performed using a Dionex RSLCnano HPLC. ..

    Article Title: Loss of NAD-Dependent Protein Deacetylase Sirtuin-2 Alters Mitochondrial Protein Acetylation and Dysregulates Mitophagy
    Article Snippet: .. Liquid chromatography-mass spectrometry analysis of the peptides was performed using an LTQ-Orbitrap mass spectrometer (Thermo Scientific) that was equipped with a nanospray source and an Eksigent NanoLC 1D Plus and AS1 Autosampler. ..

    SDS Page:

    Article Title: The function of the PduJ microcompartment shell protein is determined by the genomic position of its encoding gene
    Article Snippet: To verify the incorporation of PduJM into microcompartment shell, the MCP particles were purified and fractionated by SDS PAGE. .. The trypsin digested peptides were extracted and subjected to liquid chromatography mass spectrometry analysis according to the manufacturer's instructions (Thermo, Q Exactive™ Hybrid Quadrupole-Obrbitrap Mass Spectrometer).

    Liquid Chromatography:

    Article Title: Extraction and biomolecular analysis of dermal interstitial fluid collected with hollow microneedles
    Article Snippet: .. Liquid chromatography mass spectrometry analysis (LC-MS/MS) Each pH 10 fraction was re-suspended in 20 µl of acetonitrile/water/formic acid 5/95/0.5 (v/v/v) and 2 µl was analyzed in a nanoLC system (Dionex, Ultimate 3000) coupled to a Q Exactive plus mass spectrometer (Thermo Fisher Scientific, Waltham, MA). .. Peptides were loaded on a PepMap trap 300 um x 5 mm C18, 100 A (Thermo Fisher Scientific, Waltham, MA) at flow rate of 10 µl per min for 5 min. Peptides were then separated on an EASYspray C18 75 µm x 50 cm analytical column over a 182-min run at flow rate of 300 nl/min using mobile phase A of 0.1% formic acid in water and mobile phase B of acetonitrile/water/formic acid 80/20/0.1 (v/v/v).

    Article Title: Profiling the effect of nafcillin on HA-MRSA D712 using bacteriological and physiological media
    Article Snippet: Paragraph title: Untargeted liquid chromatography mass spectrometry data acquisition ... For LC/MS analysis, samples were subjected to chromatographic separation using an UltiMate 3000 UHPLC system (Thermo Scientific).

    Article Title: The function of the PduJ microcompartment shell protein is determined by the genomic position of its encoding gene
    Article Snippet: .. The trypsin digested peptides were extracted and subjected to liquid chromatography mass spectrometry analysis according to the manufacturer's instructions (Thermo, Q Exactive™ Hybrid Quadrupole-Obrbitrap Mass Spectrometer). .. Tandem mass spectra were searched against the known protein sequence from S. enterica serovar Typhimurium LT2 using both MASCOT and Sequest HT (Thermo Proteome discoverer software from Thermo Fisher) with maximum mixed cleavage sites 2, fixed carbamidomethylation and variable methionine oxidation.

    Article Title: Biochemical assessment of red blood cells during storage by 1H nuclear magnetic resonance spectroscopy. Identification of a biomarker of their level of protection against oxidative stress
    Article Snippet: .. Liquid chromatography mass spectrometry analysis of the NMR samples of RBC supernant and lysates was performed using a LTQ Orbitrap XL (Thermo Scientific, Waltham, MA, USA) coupled to a HPLC Dionex Ultimate 3000. .. The liquid chromatography separation was carried out on Aeris peptide 3.6u XB-C18 column (150×2.1 mm) (Phenomenex, Torrance, CA, USA).

    Article Title: A proteomic chronology of gene expression through the cell cycle in human myeloid leukemia cells
    Article Snippet: .. Liquid chromatography mass spectrometry analysis (LC-MS/MS) For tryptic digests, including tryptic + Lys-C double digests, peptide chromatography was performed using a Dionex RSLCnano HPLC. ..

    Centrifugation:

    Article Title: HLA-F Allele-Specific Peptide Restriction Represents an Exceptional Proteomic Footprint
    Article Snippet: Gel pieces were destained with 50% ACN/50mM Ammoniumcarbonat (ABC) and dehydrated with 100% ACN and dried via vacuum centrifugation. .. The LC/MS analysis was performed using a Dionex Ultimate 3000 high-performance LC system and a LTQ Orbitrap Lumos Mass Spectrometer (Thermo Fisher Scientific) in data-dependent acquisition (DDA) mode.

    Nuclear Magnetic Resonance:

    Article Title: Biochemical assessment of red blood cells during storage by 1H nuclear magnetic resonance spectroscopy. Identification of a biomarker of their level of protection against oxidative stress
    Article Snippet: .. Liquid chromatography mass spectrometry analysis of the NMR samples of RBC supernant and lysates was performed using a LTQ Orbitrap XL (Thermo Scientific, Waltham, MA, USA) coupled to a HPLC Dionex Ultimate 3000. .. The liquid chromatography separation was carried out on Aeris peptide 3.6u XB-C18 column (150×2.1 mm) (Phenomenex, Torrance, CA, USA).

    Purification:

    Article Title: The function of the PduJ microcompartment shell protein is determined by the genomic position of its encoding gene
    Article Snippet: To verify the incorporation of PduJM into microcompartment shell, the MCP particles were purified and fractionated by SDS PAGE. .. The trypsin digested peptides were extracted and subjected to liquid chromatography mass spectrometry analysis according to the manufacturer's instructions (Thermo, Q Exactive™ Hybrid Quadrupole-Obrbitrap Mass Spectrometer).

    Article Title: The corrinoid cofactor of reductive dehalogenases affects dechlorination rates and extents in organohalide-respiring Dehalococcoides mccartyi
    Article Snippet: Cobamides were detected at 361 nm with an Agilent 1260 Infinity DAD and quantified by comparing integrated peak areas with 4-point calibration curves generated with purified cobamides. .. Liquid chromatography–mass spectrometry analysis was performed using a Dionex Ultimate 3000 system (Thermo Fisher Scientific) with an inline DAD fitted to an Exactive Plus Orbitrap Mass Spectrometer with an electrospray ionization source (Thermo Fisher Scientific).

    Concentration Assay:

    Article Title: UGT85A84 Catalyzes the Glycosylation of Aromatic Monoterpenes in Osmanthus fragrans Lour. Flowers
    Article Snippet: To determine the kinetic parameters of UDP-glucose, UDP-glucose concentration was set from 25 µM to 10 mM with a constant linalool or linalool oxide concentration of 0.5 mM. .. LC-MS analysis was performed in the Thermo Scientific™ UltiMate™ 3000RS system.

    Article Title: Pharmacokinetic Study of Bioactive Glycopeptide from Strongylocentrotus droebachiensis After Intranasal Administration to Rats Using Biomarker Approach
    Article Snippet: This was followed by alkylation with 500 mM iodacetamide (IAA, Sigma, St. Louis, MI, USA) in a final concentration of 60 mM IAA performed for 30 min at room temperature. .. Before LC-MS analysis, peptide extracts were desalted and cleaned up using C18 spin tips (Pierce, Waltham, MS, USA) according to the manufacturer’s protocol.

    Generated:

    Article Title: The corrinoid cofactor of reductive dehalogenases affects dechlorination rates and extents in organohalide-respiring Dehalococcoides mccartyi
    Article Snippet: Cobamides were detected at 361 nm with an Agilent 1260 Infinity DAD and quantified by comparing integrated peak areas with 4-point calibration curves generated with purified cobamides. .. Liquid chromatography–mass spectrometry analysis was performed using a Dionex Ultimate 3000 system (Thermo Fisher Scientific) with an inline DAD fitted to an Exactive Plus Orbitrap Mass Spectrometer with an electrospray ionization source (Thermo Fisher Scientific).

    Liquid Chromatography with Mass Spectroscopy:

    Article Title: HLA-F Allele-Specific Peptide Restriction Represents an Exceptional Proteomic Footprint
    Article Snippet: .. The LC/MS analysis was performed using a Dionex Ultimate 3000 high-performance LC system and a LTQ Orbitrap Lumos Mass Spectrometer (Thermo Fisher Scientific) in data-dependent acquisition (DDA) mode. ..

    Article Title: Profiling the effect of nafcillin on HA-MRSA D712 using bacteriological and physiological media
    Article Snippet: .. For LC/MS analysis, samples were subjected to chromatographic separation using an UltiMate 3000 UHPLC system (Thermo Scientific). .. Chromatographic separations were achieved using a 50 mm × 2.1 mm Kinetex 2.6 micron polar-C18 column (Phenomenex) held at a fixed temperature of 30 °C within an actively heated column compartment.

    Article Title: ProTargetMiner as a proteome signature library of anticancer molecules for functional discovery
    Article Snippet: .. LC-MS analysis Samples were loaded with buffer A (0.1% FA in water) onto a 50 cm EASY-Spray column (75 µm internal diameter, packed with PepMap C18, 2 µm beads, 100 Å pore size; Cat#ES803) connected to the EASY-nLC 1000 (Thermo; Cat#LC120) and eluted with a buffer B (98% ACN, 0.1% FA, 2% H2 O) gradient from 2 to 35% of at a flow rate of 250 nL min−1 . .. Mass spectra were acquired with an Orbitrap Q Exactive Plus mass spectrometer (Thermo; Cat# IQLAAEGAAPFALGMBDK) in the data-dependent mode with MS1 scan at 70,000 resolution, and MS2 at 35,000, in the m/z range from 375 to 1400.

    Article Title: The corrinoid cofactor of reductive dehalogenases affects dechlorination rates and extents in organohalide-respiring Dehalococcoides mccartyi
    Article Snippet: Cobamide purity and authenticity (for example, distinct singular peaks and m / z values) were analyzed in a combined approach employing an Agilent (Santa Clara, CA, USA) 1200 HPLC system and a Thermo Fisher Scientific (Waltham, MA, USA) Orbitrap Exactive Plus LC/MS system ( ). .. Liquid chromatography–mass spectrometry analysis was performed using a Dionex Ultimate 3000 system (Thermo Fisher Scientific) with an inline DAD fitted to an Exactive Plus Orbitrap Mass Spectrometer with an electrospray ionization source (Thermo Fisher Scientific).

    Article Title: mTORC1 Activation Requires DRAM-1 by Facilitating Lysosomal Amino Acid Efflux
    Article Snippet: .. LC-MS Analysis A Q Exactive Orbitrap mass spectrometer (Thermo Scientific, Waltham, MA, USA) was used together with a Thermo Ultimate 3000 HPLC system. .. The HPLC setup consisted of a ZIC-pHILIC column (SeQuant, 150 × 2.1mm, 5 μm, Merck KGaA, Darmstadt, Germany), with a ZIC-pHILIC guard column (SeQuant, 20 × 2.1mm) and an initial mobile phase of 20% 20mM ammonium carbonate, pH 9.4, and 80% acetonitrile.

    Article Title: Choosing an Optimal Sample Preparation in Caulobacter crescentus for Untargeted Metabolomics Approaches
    Article Snippet: .. Extract Preparation for LC-MS Analysis The supernatants were collected, evaporated to dryness using a SpeedVac (ThermoFisher, Langenselbold, Germany) and reconstituted in 100 µL ACN:H2 O 50:50. ..

    Article Title: UGT85A84 Catalyzes the Glycosylation of Aromatic Monoterpenes in Osmanthus fragrans Lour. Flowers
    Article Snippet: .. LC-MS analysis was performed in the Thermo Scientific™ UltiMate™ 3000RS system. .. The separation was conducted on a Thermo Scientific™ Syncronis C18 column (100*2.1 mm, 1.7µm) at 40°C.

    Article Title: Pharmacokinetic Study of Bioactive Glycopeptide from Strongylocentrotus droebachiensis After Intranasal Administration to Rats Using Biomarker Approach
    Article Snippet: .. Before LC-MS analysis, peptide extracts were desalted and cleaned up using C18 spin tips (Pierce, Waltham, MS, USA) according to the manufacturer’s protocol. .. The flow-through peptide sample was dissolved in 15 µL 0.1% trifluoroacetic acid (TFA, Fisher Scientific, Waltham, MS, USA) and 6 µL was injected into the LC-MS system.

    Article Title: The integration of metabolome and proteome reveals bioactive polyphenols and hispidin in ARTP mutagenized Phellinus baumii
    Article Snippet: .. LC/MS analysis and statistical analysis Refering to previous research method , the ACQUITY UHPLC system Ultimate 3000 (Thermo Fisher Scientific, Waltham, MA, USA) coupled with LTQ Orbitrap MS (Thermo Fisher Scientific, Waltham, MA, USA) was used for metabolic profiling in both ESI+ and ESI− ion modes. ..

    Sequencing:

    Article Title: The function of the PduJ microcompartment shell protein is determined by the genomic position of its encoding gene
    Article Snippet: The trypsin digested peptides were extracted and subjected to liquid chromatography mass spectrometry analysis according to the manufacturer's instructions (Thermo, Q Exactive™ Hybrid Quadrupole-Obrbitrap Mass Spectrometer). .. Tandem mass spectra were searched against the known protein sequence from S. enterica serovar Typhimurium LT2 using both MASCOT and Sequest HT (Thermo Proteome discoverer software from Thermo Fisher) with maximum mixed cleavage sites 2, fixed carbamidomethylation and variable methionine oxidation.

    Staining:

    Article Title: HLA-F Allele-Specific Peptide Restriction Represents an Exceptional Proteomic Footprint
    Article Snippet: Gels were stained with Coomassi SimplyBlue™ SafeStain (Thermo Fischer). .. The LC/MS analysis was performed using a Dionex Ultimate 3000 high-performance LC system and a LTQ Orbitrap Lumos Mass Spectrometer (Thermo Fisher Scientific) in data-dependent acquisition (DDA) mode.

    Chloramphenicol Acetyltransferase Assay:

    Article Title: ProTargetMiner as a proteome signature library of anticancer molecules for functional discovery
    Article Snippet: .. LC-MS analysis Samples were loaded with buffer A (0.1% FA in water) onto a 50 cm EASY-Spray column (75 µm internal diameter, packed with PepMap C18, 2 µm beads, 100 Å pore size; CatES803) connected to the EASY-nLC 1000 (Thermo; Cat#LC120) and eluted with a buffer B (98% ACN, 0.1% FA, 2% H2 O) gradient from 2 to 35% of at a flow rate of 250 nL min−1 . .. Mass spectra were acquired with an Orbitrap Q Exactive Plus mass spectrometer (Thermo; Cat# IQLAAEGAAPFALGMBDK) in the data-dependent mode with MS1 scan at 70,000 resolution, and MS2 at 35,000, in the m/z range from 375 to 1400.

    Injection:

    Article Title: Profiling the effect of nafcillin on HA-MRSA D712 using bacteriological and physiological media
    Article Snippet: For LC/MS analysis, samples were subjected to chromatographic separation using an UltiMate 3000 UHPLC system (Thermo Scientific). .. Samples were injected onto the LC column via thermostatted autosampler maintained at 4 °C.

    Article Title: The corrinoid cofactor of reductive dehalogenases affects dechlorination rates and extents in organohalide-respiring Dehalococcoides mccartyi
    Article Snippet: The column was equilibrated with 82% eluent A/18% eluent B, and a linear change to 75% A/25% B was applied following sample injection over a 12-min time period. .. Liquid chromatography–mass spectrometry analysis was performed using a Dionex Ultimate 3000 system (Thermo Fisher Scientific) with an inline DAD fitted to an Exactive Plus Orbitrap Mass Spectrometer with an electrospray ionization source (Thermo Fisher Scientific).

    Article Title: A Resveratrol Analogue Promotes ERKMAPK
    Article Snippet: Liquid chromatography–mass spectrometry analysis was carried out on a Q-Exactive mass spectrometer coupled to an HTC autosampler and Accela pump (all from Thermo Fisher Scientific, Waltham, MA). .. Twenty-five microliters of the standard or extracts were injected onto an Ascentis Express C18 column (15 × 3 mm; 2.7 µ m; Supelco, Sigma-Aldrich).

    Article Title: mTORC1 Activation Requires DRAM-1 by Facilitating Lysosomal Amino Acid Efflux
    Article Snippet: LC-MS Analysis A Q Exactive Orbitrap mass spectrometer (Thermo Scientific, Waltham, MA, USA) was used together with a Thermo Ultimate 3000 HPLC system. .. Cell and media extracts (5 μl) were injected and metabolites were separated over a 15 minute mobile phase gradient, decreasing the acetonitrile content to 20%, at a flow rate of 200μL/min and a column temperature of 45°C.

    Article Title: Choosing an Optimal Sample Preparation in Caulobacter crescentus for Untargeted Metabolomics Approaches
    Article Snippet: Extract Preparation for LC-MS Analysis The supernatants were collected, evaporated to dryness using a SpeedVac (ThermoFisher, Langenselbold, Germany) and reconstituted in 100 µL ACN:H2 O 50:50. .. QC and dQC were injected at regular intervals throughout the LC-MS analyses to assess analytical variability.

    Article Title: Pharmacokinetic Study of Bioactive Glycopeptide from Strongylocentrotus droebachiensis After Intranasal Administration to Rats Using Biomarker Approach
    Article Snippet: Before LC-MS analysis, peptide extracts were desalted and cleaned up using C18 spin tips (Pierce, Waltham, MS, USA) according to the manufacturer’s protocol. .. The flow-through peptide sample was dissolved in 15 µL 0.1% trifluoroacetic acid (TFA, Fisher Scientific, Waltham, MS, USA) and 6 µL was injected into the LC-MS system.

    Enzymatic Assay:

    Article Title: UGT85A84 Catalyzes the Glycosylation of Aromatic Monoterpenes in Osmanthus fragrans Lour. Flowers
    Article Snippet: Paragraph title: OfUGT85A84 Enzyme Assay ... LC-MS analysis was performed in the Thermo Scientific™ UltiMate™ 3000RS system.

    Software:

    Article Title: The function of the PduJ microcompartment shell protein is determined by the genomic position of its encoding gene
    Article Snippet: The trypsin digested peptides were extracted and subjected to liquid chromatography mass spectrometry analysis according to the manufacturer's instructions (Thermo, Q Exactive™ Hybrid Quadrupole-Obrbitrap Mass Spectrometer). .. Tandem mass spectra were searched against the known protein sequence from S. enterica serovar Typhimurium LT2 using both MASCOT and Sequest HT (Thermo Proteome discoverer software from Thermo Fisher) with maximum mixed cleavage sites 2, fixed carbamidomethylation and variable methionine oxidation.

    Article Title: UGT85A84 Catalyzes the Glycosylation of Aromatic Monoterpenes in Osmanthus fragrans Lour. Flowers
    Article Snippet: The apparent K m and k cat values for each aglycone and sugar donor were determined by Michaelis-Menten fitting in OriginPro 9.1 software. .. LC-MS analysis was performed in the Thermo Scientific™ UltiMate™ 3000RS system.

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    Thermo Fisher lc ms ms analysis all peptide
    GPS leads to the identification of ADP-ribosylated ARTDs/PARPs other than ARTD1/PARP1. (A) A comparison using Venn diagrams for ADPr peptides found in two replicates for full scan (400–1500 m / z ) and combined 4× GPS scans (GPS-1, 400–605; GPS-2, 595–805; GPS-3, 795–1005; GPS-4, 995–1200 m / z ). (B) A comparison of ADPr peptides found in control and IFN-γ-treated THP-1 cells for the full scan and combined 4× GPS scans. (C) Sequence motif <t>analysis</t> for ADPr acceptor amino acids (N, number of ADPr peptides used for the analysis). (D) A plot of the number of ADP-ribosylation sites per protein. (E) Comparison of ADPr <t>peptide</t> abundances between control and IFN-γ in each replicate; regression lines, 95% confidence interval, and standard error of estimate (SEE) are provided (red dots are outliers). (F) <t>MS/MS</t> spectra of an ARTD8/PARP14 ADPr peptide using PRM acquisitions. Black peaks were manually annotated. *, ADPr site. (G) A comparison of the number of proteins identified in the Af1521 elution (ADPr proteins) and input samples (backbone proteins) per replicate. (H) A comparison of the relative changes to ADPr peptides versus their backbone proteins in response to IFN-γ (IFN-γ/control).
    Lc Ms Ms Analysis All Peptide, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 77/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Thermo Fisher lc ms analysis lc ms
    Chromatographic profile of the Impatiens glandulifera Royle methanolic extract of the roots measured as total ion current (TIC) by <t>LC-MS</t> (APCI). MS spectrum of THNG is shown on the Figure 3 .
    Lc Ms Analysis Lc Ms, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 80/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Thermo Fisher enriched proteins lc ms analysis
    Heatmap visualization of quantitative values of the 182 <t>proteins</t> <t>enriched</t> at the protein level . Left: heatmap of the 182 proteins identified and quantified over all six HNE exposure groups; Right: Zoomed-in region of the heatmap focusing on the most abundantly detected and identified putative HNE protein adducts. After affinity capture, samples were trypsin-digested and analyzed using <t>LC-MS.</t> Protein quantification was based on the peak intensity of the 3 most intense peptides. A total of 182 proteins were quantified. From top to bottom the 182 identified proteins are listed and each row represents a protein and its corresponding abundance. From left to right, HNE concentrations that were used for the in vitro exposure experiments of the mitochondrial protein samples. The color in each cell represents protein abundance obtained from the “Hi3” peptide intensity approach: red is more abundant and dark green is less abundant. Black indicates missing values.
    Enriched Proteins Lc Ms Analysis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher lc esi ms analysis
    HPLC-UV, <t>LC-ESI-HRMS</t> and MS–MS analysis of P450 deletion mutants from both Tü 3010 and <t>thiolactomycin</t> (TLM) biosynthetic pathways. (A) HPLC trace profiles (UV 238 nm ) of extracts from S. thiolactonus wild-type strain NRRL 15439 and mutants ΔstuD1 and ΔstuD2, the Lentzea sp. wild-type strain ATCC31319 and ΔtlmD1 mutant. Separation was achieved as described in the materials and methods section. Production of Tü 3010 (retention time 8.39 min) was abolished in both P450 ( stuD1 and stuD2 ) mutants, although the ΔstuD2 mutant produced a new UV-absorbing peak (retention time 10.93 min). Thiolactomycin (retention time 27.14 min) production was lost, with no obvious new UV-absorbing peak, upon disruption of tlmD1 . (B) Further LC-ESI-HRMS analysis of the ΔstuD2 intermediate by selective ion monitoring confirmed it as thiotetromycin ( 2 ) (Fig. S20 † ). Asterisk denotes not detected.
    Lc Esi Ms Analysis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    GPS leads to the identification of ADP-ribosylated ARTDs/PARPs other than ARTD1/PARP1. (A) A comparison using Venn diagrams for ADPr peptides found in two replicates for full scan (400–1500 m / z ) and combined 4× GPS scans (GPS-1, 400–605; GPS-2, 595–805; GPS-3, 795–1005; GPS-4, 995–1200 m / z ). (B) A comparison of ADPr peptides found in control and IFN-γ-treated THP-1 cells for the full scan and combined 4× GPS scans. (C) Sequence motif analysis for ADPr acceptor amino acids (N, number of ADPr peptides used for the analysis). (D) A plot of the number of ADP-ribosylation sites per protein. (E) Comparison of ADPr peptide abundances between control and IFN-γ in each replicate; regression lines, 95% confidence interval, and standard error of estimate (SEE) are provided (red dots are outliers). (F) MS/MS spectra of an ARTD8/PARP14 ADPr peptide using PRM acquisitions. Black peaks were manually annotated. *, ADPr site. (G) A comparison of the number of proteins identified in the Af1521 elution (ADPr proteins) and input samples (backbone proteins) per replicate. (H) A comparison of the relative changes to ADPr peptides versus their backbone proteins in response to IFN-γ (IFN-γ/control).

    Journal: Journal of Proteome Research

    Article Title: A Study into the ADP-Ribosylome of IFN-γ-Stimulated THP-1 Human Macrophage-like Cells Identifies ARTD8/PARP14 and ARTD9/PARP9 ADP-Ribosylation

    doi: 10.1021/acs.jproteome.8b00895

    Figure Lengend Snippet: GPS leads to the identification of ADP-ribosylated ARTDs/PARPs other than ARTD1/PARP1. (A) A comparison using Venn diagrams for ADPr peptides found in two replicates for full scan (400–1500 m / z ) and combined 4× GPS scans (GPS-1, 400–605; GPS-2, 595–805; GPS-3, 795–1005; GPS-4, 995–1200 m / z ). (B) A comparison of ADPr peptides found in control and IFN-γ-treated THP-1 cells for the full scan and combined 4× GPS scans. (C) Sequence motif analysis for ADPr acceptor amino acids (N, number of ADPr peptides used for the analysis). (D) A plot of the number of ADP-ribosylation sites per protein. (E) Comparison of ADPr peptide abundances between control and IFN-γ in each replicate; regression lines, 95% confidence interval, and standard error of estimate (SEE) are provided (red dots are outliers). (F) MS/MS spectra of an ARTD8/PARP14 ADPr peptide using PRM acquisitions. Black peaks were manually annotated. *, ADPr site. (G) A comparison of the number of proteins identified in the Af1521 elution (ADPr proteins) and input samples (backbone proteins) per replicate. (H) A comparison of the relative changes to ADPr peptides versus their backbone proteins in response to IFN-γ (IFN-γ/control).

    Article Snippet: LC–MS/MS Analysis All peptide samples were analyzed on an Orbitrap Fusion Lumos mass spectrometer fronted with an EASY-Spray Source, coupled to an Easy-nLC1000 HPLC pump (Thermo Fisher Scientific).

    Techniques: Sequencing, Mass Spectrometry

    Data processing of product ion triggered MS/MS spectra. (A) A schematic of SEQUEST-HT searches of triggered EThcD and HCD spectra using the second Af1521 replicate of IFN-γ-treated THP-1 cells. (B) Number of peptide-spectrum matches (PSMs) of assigned ADPr and unmodified peptides from the triggered spectra. (C–E) Distribution of isolation interference for product ion triggered or DDA PSMs. (F) Number of ADPr peptides with high confidence detected by either EThcD or HCD. (G) Venn diagrams comparing ADPr peptide identifications between EThcD and HCD for all ADPr peptides, and those with > 95% ADPr acceptor site probability.

    Journal: Journal of Proteome Research

    Article Title: A Study into the ADP-Ribosylome of IFN-γ-Stimulated THP-1 Human Macrophage-like Cells Identifies ARTD8/PARP14 and ARTD9/PARP9 ADP-Ribosylation

    doi: 10.1021/acs.jproteome.8b00895

    Figure Lengend Snippet: Data processing of product ion triggered MS/MS spectra. (A) A schematic of SEQUEST-HT searches of triggered EThcD and HCD spectra using the second Af1521 replicate of IFN-γ-treated THP-1 cells. (B) Number of peptide-spectrum matches (PSMs) of assigned ADPr and unmodified peptides from the triggered spectra. (C–E) Distribution of isolation interference for product ion triggered or DDA PSMs. (F) Number of ADPr peptides with high confidence detected by either EThcD or HCD. (G) Venn diagrams comparing ADPr peptide identifications between EThcD and HCD for all ADPr peptides, and those with > 95% ADPr acceptor site probability.

    Article Snippet: LC–MS/MS Analysis All peptide samples were analyzed on an Orbitrap Fusion Lumos mass spectrometer fronted with an EASY-Spray Source, coupled to an Easy-nLC1000 HPLC pump (Thermo Fisher Scientific).

    Techniques: Mass Spectrometry, Isolation

    Chromatographic profile of the Impatiens glandulifera Royle methanolic extract of the roots measured as total ion current (TIC) by LC-MS (APCI). MS spectrum of THNG is shown on the Figure 3 .

    Journal: Molecules

    Article Title: Separation and Identification of 1,2,4-Trihydroxynaphthalene-1-O-glucoside in Impatiens glandulifera Royle

    doi: 10.3390/molecules18078429

    Figure Lengend Snippet: Chromatographic profile of the Impatiens glandulifera Royle methanolic extract of the roots measured as total ion current (TIC) by LC-MS (APCI). MS spectrum of THNG is shown on the Figure 3 .

    Article Snippet: LC-MS Analysis LC-MS was performed using LCQ Accela Fleet (Thermo Fisher Scientific, San Jose, CA, USA) with atmospheric pressure chemical (APCI) and a photodiode array detector.

    Techniques: Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry

    Heatmap visualization of quantitative values of the 182 proteins enriched at the protein level . Left: heatmap of the 182 proteins identified and quantified over all six HNE exposure groups; Right: Zoomed-in region of the heatmap focusing on the most abundantly detected and identified putative HNE protein adducts. After affinity capture, samples were trypsin-digested and analyzed using LC-MS. Protein quantification was based on the peak intensity of the 3 most intense peptides. A total of 182 proteins were quantified. From top to bottom the 182 identified proteins are listed and each row represents a protein and its corresponding abundance. From left to right, HNE concentrations that were used for the in vitro exposure experiments of the mitochondrial protein samples. The color in each cell represents protein abundance obtained from the “Hi3” peptide intensity approach: red is more abundant and dark green is less abundant. Black indicates missing values.

    Journal: Frontiers in Chemistry

    Article Title: Label-Free Proteomics Assisted by Affinity Enrichment for Elucidating the Chemical Reactivity of the Liver Mitochondrial Proteome toward Adduction by the Lipid Electrophile 4-hydroxy-2-nonenal (HNE)

    doi: 10.3389/fchem.2016.00002

    Figure Lengend Snippet: Heatmap visualization of quantitative values of the 182 proteins enriched at the protein level . Left: heatmap of the 182 proteins identified and quantified over all six HNE exposure groups; Right: Zoomed-in region of the heatmap focusing on the most abundantly detected and identified putative HNE protein adducts. After affinity capture, samples were trypsin-digested and analyzed using LC-MS. Protein quantification was based on the peak intensity of the 3 most intense peptides. A total of 182 proteins were quantified. From top to bottom the 182 identified proteins are listed and each row represents a protein and its corresponding abundance. From left to right, HNE concentrations that were used for the in vitro exposure experiments of the mitochondrial protein samples. The color in each cell represents protein abundance obtained from the “Hi3” peptide intensity approach: red is more abundant and dark green is less abundant. Black indicates missing values.

    Article Snippet: LC-MS-based quantitative analysis of enriched proteins LC-MS analysis of the peptide samples from protein level or peptide level enrichment was performed using a hybrid linear ion trap-Fourier transform ion cyclotron resonance mass spectrometer (LTQ-FT Ultra, Thermo Fisher) coupled to a NanoAcquity UPLC (Waters, MA) equipped with a BEH C18 column (100 μm × 15 cm) (Waters Corp.).

    Techniques: Liquid Chromatography with Mass Spectroscopy, In Vitro

    HPLC-UV, LC-ESI-HRMS and MS–MS analysis of P450 deletion mutants from both Tü 3010 and thiolactomycin (TLM) biosynthetic pathways. (A) HPLC trace profiles (UV 238 nm ) of extracts from S. thiolactonus wild-type strain NRRL 15439 and mutants ΔstuD1 and ΔstuD2, the Lentzea sp. wild-type strain ATCC31319 and ΔtlmD1 mutant. Separation was achieved as described in the materials and methods section. Production of Tü 3010 (retention time 8.39 min) was abolished in both P450 ( stuD1 and stuD2 ) mutants, although the ΔstuD2 mutant produced a new UV-absorbing peak (retention time 10.93 min). Thiolactomycin (retention time 27.14 min) production was lost, with no obvious new UV-absorbing peak, upon disruption of tlmD1 . (B) Further LC-ESI-HRMS analysis of the ΔstuD2 intermediate by selective ion monitoring confirmed it as thiotetromycin ( 2 ) (Fig. S20 † ). Asterisk denotes not detected.

    Journal: Chemical Science

    Article Title: A genomics-led approach to deciphering the mechanism of thiotetronate antibiotic biosynthesis genomics-led approach to deciphering the mechanism of thiotetronate antibiotic biosynthesis †Electronic supplementary information (ESI) available: Fig. S1–S21; Tables S1–S5, full experimental details and procedures. See DOI: 10.1039/c5sc03059eClick here for additional data file.

    doi: 10.1039/c5sc03059e

    Figure Lengend Snippet: HPLC-UV, LC-ESI-HRMS and MS–MS analysis of P450 deletion mutants from both Tü 3010 and thiolactomycin (TLM) biosynthetic pathways. (A) HPLC trace profiles (UV 238 nm ) of extracts from S. thiolactonus wild-type strain NRRL 15439 and mutants ΔstuD1 and ΔstuD2, the Lentzea sp. wild-type strain ATCC31319 and ΔtlmD1 mutant. Separation was achieved as described in the materials and methods section. Production of Tü 3010 (retention time 8.39 min) was abolished in both P450 ( stuD1 and stuD2 ) mutants, although the ΔstuD2 mutant produced a new UV-absorbing peak (retention time 10.93 min). Thiolactomycin (retention time 27.14 min) production was lost, with no obvious new UV-absorbing peak, upon disruption of tlmD1 . (B) Further LC-ESI-HRMS analysis of the ΔstuD2 intermediate by selective ion monitoring confirmed it as thiotetromycin ( 2 ) (Fig. S20 † ). Asterisk denotes not detected.

    Article Snippet: LC-ESI-MS analysis of thiolactomycin was performed on a Finnigan LTQ mass spectrometer (Thermo Scientific), operating in positive ion mode.

    Techniques: High Performance Liquid Chromatography, Mass Spectrometry, Mutagenesis, Produced