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    Structured Review

    Thermo Fisher nano hplc
    Experimental design of the quantitative proteomics of Brucella abortus under environmental stress. A label-free relative quantitative proteomics approach was utilized to investigate and compare the global proteomic changes of B. abortus in response to a variety of distinct stresses including a control condition, seven single-stress conditions, and a multi-stress condition. Protein samples were prepared and proteolytic-digested using trypsin enzymes. Peptides were analyzed on a Q-Exactive HF MS coupled online with a <t>nano-HPLC.</t> The identified proteins were quantified using a label-free approach and further functionally analyzed using the COG and KEGG databases.
    Nano Hplc, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 132 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nano hplc/product/Thermo Fisher
    Average 99 stars, based on 132 article reviews
    Price from $9.99 to $1999.99
    nano hplc - by Bioz Stars, 2020-04
    99/100 stars

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    1) Product Images from "Relative Quantitative Proteomic Analysis of Brucella abortus Reveals Metabolic Adaptation to Multiple Environmental Stresses"

    Article Title: Relative Quantitative Proteomic Analysis of Brucella abortus Reveals Metabolic Adaptation to Multiple Environmental Stresses

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2017.02347

    Experimental design of the quantitative proteomics of Brucella abortus under environmental stress. A label-free relative quantitative proteomics approach was utilized to investigate and compare the global proteomic changes of B. abortus in response to a variety of distinct stresses including a control condition, seven single-stress conditions, and a multi-stress condition. Protein samples were prepared and proteolytic-digested using trypsin enzymes. Peptides were analyzed on a Q-Exactive HF MS coupled online with a nano-HPLC. The identified proteins were quantified using a label-free approach and further functionally analyzed using the COG and KEGG databases.
    Figure Legend Snippet: Experimental design of the quantitative proteomics of Brucella abortus under environmental stress. A label-free relative quantitative proteomics approach was utilized to investigate and compare the global proteomic changes of B. abortus in response to a variety of distinct stresses including a control condition, seven single-stress conditions, and a multi-stress condition. Protein samples were prepared and proteolytic-digested using trypsin enzymes. Peptides were analyzed on a Q-Exactive HF MS coupled online with a nano-HPLC. The identified proteins were quantified using a label-free approach and further functionally analyzed using the COG and KEGG databases.

    Techniques Used: Mass Spectrometry, High Performance Liquid Chromatography

    Related Articles

    Centrifugation:

    Article Title: Oligoribonuclease is a common downstream target of lithium-induced pAp accumulation in Escherichia coli and human cells
    Article Snippet: Elution was done by adding 75 µl of hot SDS-sample buffer, and incubation at 65°C for 15 min. Beads were removed by centrifugation, and 20 µl of each sample was analyzed by gel electrophoresis on 12% SDS–PAA gels. .. Bands corresponding to proteins binding specifically to pAp were localized by staining with Bio-Safe colloidal coomassie (Bio-Rad), cut out and digested in-gel by trypsin after reduction and alkylation according to standard protocols and analyzed by nano-HPLC (LC Packing) directly coupled to an ion trap mass spectrometer (ThermoFinnigan LCQ Deca XP) equipped with a nanoelectrospray source (LC-MS/MS).

    Incubation:

    Article Title: Oligoribonuclease is a common downstream target of lithium-induced pAp accumulation in Escherichia coli and human cells
    Article Snippet: Elution was done by adding 75 µl of hot SDS-sample buffer, and incubation at 65°C for 15 min. Beads were removed by centrifugation, and 20 µl of each sample was analyzed by gel electrophoresis on 12% SDS–PAA gels. .. Bands corresponding to proteins binding specifically to pAp were localized by staining with Bio-Safe colloidal coomassie (Bio-Rad), cut out and digested in-gel by trypsin after reduction and alkylation according to standard protocols and analyzed by nano-HPLC (LC Packing) directly coupled to an ion trap mass spectrometer (ThermoFinnigan LCQ Deca XP) equipped with a nanoelectrospray source (LC-MS/MS).

    Article Title: Mouse Antibody of IgM Class is Prone to Non-Enzymatic Cleavage between CH1 and CH2 Domains
    Article Snippet: Gel pieces were re-swelled in 50 mM ABC containing 20 U of PNGase F (Promega) and incubated at 37 °C overnight. .. Then, trifluoroacetic acid (TFA) was added, peptides were collected, vacuum-dried and suspended in loading buffer (2% ACN, 0.05% TFA) for LC-MS/MS analysis done with Q Exactive mass spectrometer (Thermo Scientific) coupled with nano-HPLC (UltiMate 3000 RSLCnano System, Thermo Scientific) through a Digital PicoView 550 ion source (New Objective).

    Article Title: The EAL-domain protein FcsR regulates flagella, chemotaxis and type III secretion system in Pseudomonas aeruginosa by a phosphodiesterase independent mechanism
    Article Snippet: In-gel Cys alkylation was performed by incubation with 10 mM dithiothreitol for 1 h at 56 °C followed by incubation with 55 mM iodoacetamide at room temperature for 45 min prior to in-gel digestion. .. Tryptic peptides were separated using a nano-HPLC (EASY-nLC 1000, Thermo Scientific) coupled to an LTQ Velos mass spectrometer (Thermo Scientific).

    Article Title: Identification of potential mitochondrial CLPXP protease interactors and substrates suggests its central role in energy metabolism
    Article Snippet: After 1:1 dilution and addition of CaCl2 to a final concentration of 1 mM, the samples were incubated with trypsin (final concentration: 10 μg ml−1 ; Serva) at 37 °C overnight. .. Proteolytic digests were dissolved in 5% acetonitrile with 0.1% formic acid and loaded on reverse phase columns (trapping column: particle size 3 μm, C18, L = 20 mm; analytical column: particle size < 2 μm, C18, L = 50 cm; PepMap, Dionex/Thermo Fisher Scientific) using a nano-HPLC (Dionex – UltiMate 3000 RSLCnano, Thermo Fisher Scientific).

    Mass Spectrometry:

    Article Title: Buserelin alleviates chloride transport defect in human cystic fibrosis nasal epithelial cells
    Article Snippet: .. Nano HPLC (Dionex RSLC UltiMate3000) was used, followed by MS (Scan range: 400–1500 m/z, fragmentation: CID, Energy of collision: 35%, Cycle for fragmentation (TopN): Top8). .. Data were converted to mzXML using MS convert (ProteoWizard v 3.0.8934).

    Article Title: Relative Quantitative Proteomic Analysis of Brucella abortus Reveals Metabolic Adaptation to Multiple Environmental Stresses
    Article Snippet: .. Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) All experiments were performed on an LTQ Q-Exactive HF mass spectrometer (Thermo Scientific, USA) coupled online with a nano-HPLC (Ultimate 3000, Thermo Scientific) (Scheltema et al., ). .. The peptides were loaded onto a trap column (C18, 3 μm particles, 100 μm × 2 cm) and separated on EASY-Spray columns (C18, 1.9 μm particles, 15 μm × 12 cm) with trapping at a flow rate of 600 nL/min (Kentache et al., ).

    Article Title: Effect of Noncanonical Amino Acids on Protein–Carbohydrate Interactions: Structure, Dynamics, and Carbohydrate Affinity of a Lectin Engineered with Fluorinated Tryptophan Analogs
    Article Snippet: Possible protein variations were separated by nano-HPLC (Dionex Ultimate 3000) equipped with a Pepswift precolumn (monolithic, 5 × 0.2 mm2 ) and a ProSwift RP-4H column (monolithic, 100 μm × 25 cm) (all Thermo Fisher Scientific, Vienna, Austria). .. The maXis II ETD mass spectrometer (Bruker, Bremen, Germany) was operated with the captive spray source in positive mode with following settings: mass range, 300–3000 m /z ; 1 Hz; source voltage, 1.6 kV; dry gas flow, 3 L min–1 ; 180 °C.

    Article Title: Oligoribonuclease is a common downstream target of lithium-induced pAp accumulation in Escherichia coli and human cells
    Article Snippet: .. Bands corresponding to proteins binding specifically to pAp were localized by staining with Bio-Safe colloidal coomassie (Bio-Rad), cut out and digested in-gel by trypsin after reduction and alkylation according to standard protocols and analyzed by nano-HPLC (LC Packing) directly coupled to an ion trap mass spectrometer (ThermoFinnigan LCQ Deca XP) equipped with a nanoelectrospray source (LC-MS/MS). .. Database searches and evaluation of the results was done using the Bioworks 3.1 software.

    Article Title: Mouse Antibody of IgM Class is Prone to Non-Enzymatic Cleavage between CH1 and CH2 Domains
    Article Snippet: .. Then, trifluoroacetic acid (TFA) was added, peptides were collected, vacuum-dried and suspended in loading buffer (2% ACN, 0.05% TFA) for LC-MS/MS analysis done with Q Exactive mass spectrometer (Thermo Scientific) coupled with nano-HPLC (UltiMate 3000 RSLCnano System, Thermo Scientific) through a Digital PicoView 550 ion source (New Objective). .. Peptides were loaded onto trap column (AcclaimPepMap100 C18, Thermo Scientific; ID 75 μm, length 20 mm, particle size 3 μm, pore size 100 Å) in 2% ACN/0.05% TFA at a flow rate of 5 μl/min and then separated on analytical column (AcclaimPepMapRLSC C18, Thermo Scientific; ID 75 μm, length 150 mm, particle size 2 μm, pore size 100 Å) using a 90 min gradient of ACN from 2% to 40% in the presence of 0.05% formic acid at a flow rate of 300 nl/min.

    Article Title: MEIS homeodomain proteins facilitate PARP1/ARTD1-mediated eviction of histone H1
    Article Snippet: .. Liquid chromatography tandem mass spectrometry analyses The proteolytic digests were loaded using a nano-HPLC (Dionex RSLCnano) on reverse-phase columns (trapping column: Acclaim PepMap c18, particle size 2 µm, L = 20 mm; analytical column: Acclaim PepMap c18, particle size 2 µm, L = 25 cm; Thermo Fisher Scientific) and eluted in organic phase gradients (Buffer A: 95% H2 O, 5% DMSO, and 0.1% formic acid; Buffer B: 80% acetonitrile, 15% H2 O, 5% DMSO, and 0.1% formic acid). ..

    Article Title: Comparative proteomics of paired vocal fold and oral mucosa fibroblasts
    Article Snippet: 500 ng protein digest of each sample yielding similar total ion chromatograms (cf. in the SI) were injected and analyzed by nano-HPLC (Dionex Ultimate 3000) equipped with a C18, 5 μm, 100 Å, 5 × 0.3 mm, enrichment column and an Acclaim PepMap RSLC nanocolumn (C18, 2 μm, 100 Å, 500 × 0.075 mm) (all Thermo Fisher Scientific). .. The maXis II ETD mass spectrometer (Bruker, Vienna, Austria) was operated with the captive source in positive mode with following settings: peptide fragmentation: CID, mass range: 200–2000 m /z , 2 Hz, capillary 1300 V, dry gas flow 3 L/min with 150 °C, nanoBooster 0.2 bar, precursor acquisition control top17.

    Article Title: Proteome Analysis for Understanding Abiotic Stress (Salinity and Drought) Tolerance in Date Palm (Phoenix dactylifera L.)
    Article Snippet: .. The resulting peptides were analyzed by nano-HPLC (UltiMate 3000 HPLC System, LC Packings, Dionex, Idstein, Germany) coupled to an amaZon ETD MS ion trap spectrometer (Bruker Daltonics, Bremen, Germany) using nano-ESI spray. .. The nano-HPLC system and the ion trap spectrometer were controlled using the Bruker Compass HyStar v3.2-SR2 software.

    Article Title: Mechanisms of Yersinia YopO kinase substrate specificity
    Article Snippet: LC/MS analysis for in vitro phosphorylation Vacuum dried samples were reconstituted in 0.1% formic acid and analyzed using a nano-HPLC attached to an LTQ Orbitrap classic (Thermo Fisher Scientific). .. Survey full scan MS spectra (m/z 310–1400) were acquired with a resolution of r = 60,000 at m/z 400, an AGC target of 1e , and a maximum injection time of 1000 ms.

    Article Title: Yersinia effector protein (YopO)-mediated phosphorylation of host gelsolin causes calcium-independent activation leading to disruption of actin dynamics
    Article Snippet: Vacuum-dried samples were reconstituted in 0.1% formic acid and analyzed using a nano-HPLC coupled to an LTQ Orbitrap classic (Thermo Fisher Scientific). .. Survey full scan MS spectra (m /z 310–1400) were acquired with a resolution of r = 60,000 at m /z 400, an automatic gain control (AGC) target of 1e6 , and a maximum injection time of 1000 ms.

    Article Title: The EAL-domain protein FcsR regulates flagella, chemotaxis and type III secretion system in Pseudomonas aeruginosa by a phosphodiesterase independent mechanism
    Article Snippet: .. Tryptic peptides were separated using a nano-HPLC (EASY-nLC 1000, Thermo Scientific) coupled to an LTQ Velos mass spectrometer (Thermo Scientific). .. Peptide mixtures were injected into an Acclaim® PepMap C18 nano-trap column (75 μm × 2 cm, Thermo Scientific) and separated on a 50 μm × 150 mm C18 Easy spray column (PepMap® RSLC, 2 μm, 100 Ǻ) at a flow rate of 250 nL/min.

    Article Title: Time- and polarity-dependent proteomic changes associated with homeostatic scaling at central synapses
    Article Snippet: The dried peptide fractions were dissolved in 5% acetonitrile with 0.1% formic acid, and subsequently loaded using a nano-HPLC (Dionex U3000 RSLCnano) onto a PepMap100 trapping column (C18 , particle size 3 µm, L = 20 mm). .. Peptides eluting from the column were ionized online using a Thermo nanoFlex ESI source and analyzed in a ‘Q Exactive Plus’ mass spectrometer (Thermo Fisher Scientific).

    High Performance Liquid Chromatography:

    Article Title: Buserelin alleviates chloride transport defect in human cystic fibrosis nasal epithelial cells
    Article Snippet: .. Nano HPLC (Dionex RSLC UltiMate3000) was used, followed by MS (Scan range: 400–1500 m/z, fragmentation: CID, Energy of collision: 35%, Cycle for fragmentation (TopN): Top8). .. Data were converted to mzXML using MS convert (ProteoWizard v 3.0.8934).

    Article Title: Proteome Analysis for Understanding Abiotic Stress (Salinity and Drought) Tolerance in Date Palm (Phoenix dactylifera L.)
    Article Snippet: .. The resulting peptides were analyzed by nano-HPLC (UltiMate 3000 HPLC System, LC Packings, Dionex, Idstein, Germany) coupled to an amaZon ETD MS ion trap spectrometer (Bruker Daltonics, Bremen, Germany) using nano-ESI spray. .. The nano-HPLC system and the ion trap spectrometer were controlled using the Bruker Compass HyStar v3.2-SR2 software.

    Flow Cytometry:

    Article Title: Relative Quantitative Proteomic Analysis of Brucella abortus Reveals Metabolic Adaptation to Multiple Environmental Stresses
    Article Snippet: Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) All experiments were performed on an LTQ Q-Exactive HF mass spectrometer (Thermo Scientific, USA) coupled online with a nano-HPLC (Ultimate 3000, Thermo Scientific) (Scheltema et al., ). .. The peptides were loaded onto a trap column (C18, 3 μm particles, 100 μm × 2 cm) and separated on EASY-Spray columns (C18, 1.9 μm particles, 15 μm × 12 cm) with trapping at a flow rate of 600 nL/min (Kentache et al., ).

    Article Title: Effect of Noncanonical Amino Acids on Protein–Carbohydrate Interactions: Structure, Dynamics, and Carbohydrate Affinity of a Lectin Engineered with Fluorinated Tryptophan Analogs
    Article Snippet: Possible protein variations were separated by nano-HPLC (Dionex Ultimate 3000) equipped with a Pepswift precolumn (monolithic, 5 × 0.2 mm2 ) and a ProSwift RP-4H column (monolithic, 100 μm × 25 cm) (all Thermo Fisher Scientific, Vienna, Austria). .. One microliter of protein sample was injected and concentrated on the enrichment column for 2 min at a flow rate of 5 μL min–1 with 0.1% formic acid as isocratic solvent.

    Article Title: Mouse Antibody of IgM Class is Prone to Non-Enzymatic Cleavage between CH1 and CH2 Domains
    Article Snippet: Then, trifluoroacetic acid (TFA) was added, peptides were collected, vacuum-dried and suspended in loading buffer (2% ACN, 0.05% TFA) for LC-MS/MS analysis done with Q Exactive mass spectrometer (Thermo Scientific) coupled with nano-HPLC (UltiMate 3000 RSLCnano System, Thermo Scientific) through a Digital PicoView 550 ion source (New Objective). .. Peptides were loaded onto trap column (AcclaimPepMap100 C18, Thermo Scientific; ID 75 μm, length 20 mm, particle size 3 μm, pore size 100 Å) in 2% ACN/0.05% TFA at a flow rate of 5 μl/min and then separated on analytical column (AcclaimPepMapRLSC C18, Thermo Scientific; ID 75 μm, length 150 mm, particle size 2 μm, pore size 100 Å) using a 90 min gradient of ACN from 2% to 40% in the presence of 0.05% formic acid at a flow rate of 300 nl/min.

    Article Title: Comparative proteomics of paired vocal fold and oral mucosa fibroblasts
    Article Snippet: 500 ng protein digest of each sample yielding similar total ion chromatograms (cf. in the SI) were injected and analyzed by nano-HPLC (Dionex Ultimate 3000) equipped with a C18, 5 μm, 100 Å, 5 × 0.3 mm, enrichment column and an Acclaim PepMap RSLC nanocolumn (C18, 2 μm, 100 Å, 500 × 0.075 mm) (all Thermo Fisher Scientific). .. Samples were concentrated on the enrichment column for 2 min at a flow rate of 5 μL/min with 0.1% heptafluorobutyric acid as isocratic solvent.

    Article Title: Proteome Analysis for Understanding Abiotic Stress (Salinity and Drought) Tolerance in Date Palm (Phoenix dactylifera L.)
    Article Snippet: The resulting peptides were analyzed by nano-HPLC (UltiMate 3000 HPLC System, LC Packings, Dionex, Idstein, Germany) coupled to an amaZon ETD MS ion trap spectrometer (Bruker Daltonics, Bremen, Germany) using nano-ESI spray. .. The peptides were separated using a 45 min linear gradient from 96% (v/v) solution A (2% (v/v) acetonitrile, 0.1% (v/v) formic acid in high purity water) and 4% (v/v) solution B (98% (v/v) acetonitrile, 0.1% (v/v) formic acid in high purity water) to 50% (v/v) solution A and 50% (v/v) solution B at a flow rate of 300 nL/min.

    Article Title: The EAL-domain protein FcsR regulates flagella, chemotaxis and type III secretion system in Pseudomonas aeruginosa by a phosphodiesterase independent mechanism
    Article Snippet: Tryptic peptides were separated using a nano-HPLC (EASY-nLC 1000, Thermo Scientific) coupled to an LTQ Velos mass spectrometer (Thermo Scientific). .. Peptide mixtures were injected into an Acclaim® PepMap C18 nano-trap column (75 μm × 2 cm, Thermo Scientific) and separated on a 50 μm × 150 mm C18 Easy spray column (PepMap® RSLC, 2 μm, 100 Ǻ) at a flow rate of 250 nL/min.

    Article Title: Local and global influences on protein turnover in neurons and glia
    Article Snippet: .. LC-MS/MS Analysis The dried peptide samples were reconstituted in 5% acetonitrile with 0.1% formic acid and subsequently loaded using a nano-HPLC (Dionex U3000 RSLCnano) onto a PepMap100 loading column (C18, L = 20 mm, 3 µm particle size, Dionex, Sunnyvale, California) and washed with loading buffer (2% acetonitrile, 0.05% trifluoroacetic acid in water) for 6 min at a flow rate of 6 µL/min. .. Peptides were separated on a PepMap RSLC analytical column (C18, L = 50 cm, < 2 µm particle size, Dionex) by a gradient of phase A (water with 5% v/v dimethylsulfoxide and 0.1% formic acid) and phase B (5% dimethylsulfoxide, 15% water and 80% acetonitrile v/v/v).

    Peptide Mass Fingerprinting:

    Article Title: Identification of potential mitochondrial CLPXP protease interactors and substrates suggests its central role in energy metabolism
    Article Snippet: Paragraph title: Peptide mass fingerprinting (PMF) ... Proteolytic digests were dissolved in 5% acetonitrile with 0.1% formic acid and loaded on reverse phase columns (trapping column: particle size 3 μm, C18, L = 20 mm; analytical column: particle size < 2 μm, C18, L = 50 cm; PepMap, Dionex/Thermo Fisher Scientific) using a nano-HPLC (Dionex – UltiMate 3000 RSLCnano, Thermo Fisher Scientific).

    Concentration Assay:

    Article Title: Comparative proteomics of paired vocal fold and oral mucosa fibroblasts
    Article Snippet: Samples were acidified with formic acid (final concentration of 0.1%). .. 500 ng protein digest of each sample yielding similar total ion chromatograms (cf. in the SI) were injected and analyzed by nano-HPLC (Dionex Ultimate 3000) equipped with a C18, 5 μm, 100 Å, 5 × 0.3 mm, enrichment column and an Acclaim PepMap RSLC nanocolumn (C18, 2 μm, 100 Å, 500 × 0.075 mm) (all Thermo Fisher Scientific).

    Article Title: Identification of potential mitochondrial CLPXP protease interactors and substrates suggests its central role in energy metabolism
    Article Snippet: After 1:1 dilution and addition of CaCl2 to a final concentration of 1 mM, the samples were incubated with trypsin (final concentration: 10 μg ml−1 ; Serva) at 37 °C overnight. .. Proteolytic digests were dissolved in 5% acetonitrile with 0.1% formic acid and loaded on reverse phase columns (trapping column: particle size 3 μm, C18, L = 20 mm; analytical column: particle size < 2 μm, C18, L = 50 cm; PepMap, Dionex/Thermo Fisher Scientific) using a nano-HPLC (Dionex – UltiMate 3000 RSLCnano, Thermo Fisher Scientific).

    SDS Page:

    Article Title: Comparative proteomics of paired vocal fold and oral mucosa fibroblasts
    Article Snippet: Protein patterns a were visualized by SDS-PAGE on 4–12% NuPAGE Bis-Tris gels (Thermo Fisher Scientific), Krypton (Thermo Fisher Scientific) staining and fluorescence laser scanning on a Bio-Rad FX Pro Plus Imager. .. 500 ng protein digest of each sample yielding similar total ion chromatograms (cf. in the SI) were injected and analyzed by nano-HPLC (Dionex Ultimate 3000) equipped with a C18, 5 μm, 100 Å, 5 × 0.3 mm, enrichment column and an Acclaim PepMap RSLC nanocolumn (C18, 2 μm, 100 Å, 500 × 0.075 mm) (all Thermo Fisher Scientific).

    Tandem Mass Spectroscopy:

    Article Title: β3GnT8 Promotes Gastric Cancer Invasion by Regulating the Glycosylation of CD147
    Article Snippet: Digested peptides were analyzed by nano-HPLC (Ultimate 3000, Dionex) coupled to a linear quadrupole ion trap-Orbitrap (LTQ Orbitrap XL)mass spectrometer (Thermo Fisher) equipped with a nano-ESI source . .. All MS/MS data were analyzed using Mascot (Matrix Science, London, UK; version 2.3.01) and searched against the latest NCBI nr protein database .

    Article Title: The EAL-domain protein FcsR regulates flagella, chemotaxis and type III secretion system in Pseudomonas aeruginosa by a phosphodiesterase independent mechanism
    Article Snippet: Tryptic peptides were separated using a nano-HPLC (EASY-nLC 1000, Thermo Scientific) coupled to an LTQ Velos mass spectrometer (Thermo Scientific). .. Online MS analysis was carried out in a data dependent mode (full scan followed by MS/MS of the top 10 m/z in each segment) using a dynamic exclusion list (exclusion duration 45 s).

    Sequencing:

    Article Title: MEIS homeodomain proteins facilitate PARP1/ARTD1-mediated eviction of histone H1
    Article Snippet: Liquid chromatography tandem mass spectrometry analyses The proteolytic digests were loaded using a nano-HPLC (Dionex RSLCnano) on reverse-phase columns (trapping column: Acclaim PepMap c18, particle size 2 µm, L = 20 mm; analytical column: Acclaim PepMap c18, particle size 2 µm, L = 25 cm; Thermo Fisher Scientific) and eluted in organic phase gradients (Buffer A: 95% H2 O, 5% DMSO, and 0.1% formic acid; Buffer B: 80% acetonitrile, 15% H2 O, 5% DMSO, and 0.1% formic acid). .. Mass spectra were acquired over the m/z range 350–1,600 at a resolution of 120,000, and sequence information was acquired by computer-controlled, data-dependent automated switching to tandem mass spectrometry mode using collision energies based on mass and charge state of the candidate ions (FTIT and TOP15).

    Injection:

    Article Title: Effect of Noncanonical Amino Acids on Protein–Carbohydrate Interactions: Structure, Dynamics, and Carbohydrate Affinity of a Lectin Engineered with Fluorinated Tryptophan Analogs
    Article Snippet: Possible protein variations were separated by nano-HPLC (Dionex Ultimate 3000) equipped with a Pepswift precolumn (monolithic, 5 × 0.2 mm2 ) and a ProSwift RP-4H column (monolithic, 100 μm × 25 cm) (all Thermo Fisher Scientific, Vienna, Austria). .. One microliter of protein sample was injected and concentrated on the enrichment column for 2 min at a flow rate of 5 μL min–1 with 0.1% formic acid as isocratic solvent.

    Article Title: Comparative proteomics of paired vocal fold and oral mucosa fibroblasts
    Article Snippet: .. 500 ng protein digest of each sample yielding similar total ion chromatograms (cf. in the SI) were injected and analyzed by nano-HPLC (Dionex Ultimate 3000) equipped with a C18, 5 μm, 100 Å, 5 × 0.3 mm, enrichment column and an Acclaim PepMap RSLC nanocolumn (C18, 2 μm, 100 Å, 500 × 0.075 mm) (all Thermo Fisher Scientific). .. Samples were concentrated on the enrichment column for 2 min at a flow rate of 5 μL/min with 0.1% heptafluorobutyric acid as isocratic solvent.

    Article Title: Mechanisms of Yersinia YopO kinase substrate specificity
    Article Snippet: LC/MS analysis for in vitro phosphorylation Vacuum dried samples were reconstituted in 0.1% formic acid and analyzed using a nano-HPLC attached to an LTQ Orbitrap classic (Thermo Fisher Scientific). .. Survey full scan MS spectra (m/z 310–1400) were acquired with a resolution of r = 60,000 at m/z 400, an AGC target of 1e , and a maximum injection time of 1000 ms.

    Article Title: Yersinia effector protein (YopO)-mediated phosphorylation of host gelsolin causes calcium-independent activation leading to disruption of actin dynamics
    Article Snippet: Vacuum-dried samples were reconstituted in 0.1% formic acid and analyzed using a nano-HPLC coupled to an LTQ Orbitrap classic (Thermo Fisher Scientific). .. Survey full scan MS spectra (m /z 310–1400) were acquired with a resolution of r = 60,000 at m /z 400, an automatic gain control (AGC) target of 1e6 , and a maximum injection time of 1000 ms.

    Article Title: The EAL-domain protein FcsR regulates flagella, chemotaxis and type III secretion system in Pseudomonas aeruginosa by a phosphodiesterase independent mechanism
    Article Snippet: Tryptic peptides were separated using a nano-HPLC (EASY-nLC 1000, Thermo Scientific) coupled to an LTQ Velos mass spectrometer (Thermo Scientific). .. Peptide mixtures were injected into an Acclaim® PepMap C18 nano-trap column (75 μm × 2 cm, Thermo Scientific) and separated on a 50 μm × 150 mm C18 Easy spray column (PepMap® RSLC, 2 μm, 100 Ǻ) at a flow rate of 250 nL/min.

    Binding Assay:

    Article Title: Oligoribonuclease is a common downstream target of lithium-induced pAp accumulation in Escherichia coli and human cells
    Article Snippet: .. Bands corresponding to proteins binding specifically to pAp were localized by staining with Bio-Safe colloidal coomassie (Bio-Rad), cut out and digested in-gel by trypsin after reduction and alkylation according to standard protocols and analyzed by nano-HPLC (LC Packing) directly coupled to an ion trap mass spectrometer (ThermoFinnigan LCQ Deca XP) equipped with a nanoelectrospray source (LC-MS/MS). .. Database searches and evaluation of the results was done using the Bioworks 3.1 software.

    Nucleic Acid Electrophoresis:

    Article Title: Oligoribonuclease is a common downstream target of lithium-induced pAp accumulation in Escherichia coli and human cells
    Article Snippet: Elution was done by adding 75 µl of hot SDS-sample buffer, and incubation at 65°C for 15 min. Beads were removed by centrifugation, and 20 µl of each sample was analyzed by gel electrophoresis on 12% SDS–PAA gels. .. Bands corresponding to proteins binding specifically to pAp were localized by staining with Bio-Safe colloidal coomassie (Bio-Rad), cut out and digested in-gel by trypsin after reduction and alkylation according to standard protocols and analyzed by nano-HPLC (LC Packing) directly coupled to an ion trap mass spectrometer (ThermoFinnigan LCQ Deca XP) equipped with a nanoelectrospray source (LC-MS/MS).

    Liquid Chromatography with Mass Spectroscopy:

    Article Title: Effect of Noncanonical Amino Acids on Protein–Carbohydrate Interactions: Structure, Dynamics, and Carbohydrate Affinity of a Lectin Engineered with Fluorinated Tryptophan Analogs
    Article Snippet: Paragraph title: Intact Protein LC-MS Analysis of RSL Species ... Possible protein variations were separated by nano-HPLC (Dionex Ultimate 3000) equipped with a Pepswift precolumn (monolithic, 5 × 0.2 mm2 ) and a ProSwift RP-4H column (monolithic, 100 μm × 25 cm) (all Thermo Fisher Scientific, Vienna, Austria).

    Article Title: Mechanisms of Yersinia YopO kinase substrate specificity
    Article Snippet: .. LC/MS analysis for in vitro phosphorylation Vacuum dried samples were reconstituted in 0.1% formic acid and analyzed using a nano-HPLC attached to an LTQ Orbitrap classic (Thermo Fisher Scientific). ..

    Article Title: Yersinia effector protein (YopO)-mediated phosphorylation of host gelsolin causes calcium-independent activation leading to disruption of actin dynamics
    Article Snippet: Paragraph title: LC/MS analysis ... Vacuum-dried samples were reconstituted in 0.1% formic acid and analyzed using a nano-HPLC coupled to an LTQ Orbitrap classic (Thermo Fisher Scientific).

    Article Title: Local and global influences on protein turnover in neurons and glia
    Article Snippet: LC-MS/MS Analysis The dried peptide samples were reconstituted in 5% acetonitrile with 0.1% formic acid and subsequently loaded using a nano-HPLC (Dionex U3000 RSLCnano) onto a PepMap100 loading column (C18, L = 20 mm, 3 µm particle size, Dionex, Sunnyvale, California) and washed with loading buffer (2% acetonitrile, 0.05% trifluoroacetic acid in water) for 6 min at a flow rate of 6 µL/min. .. All solvents were LC-MS grade and purchased from Fluka.

    Fluorescence:

    Article Title: Comparative proteomics of paired vocal fold and oral mucosa fibroblasts
    Article Snippet: Protein patterns a were visualized by SDS-PAGE on 4–12% NuPAGE Bis-Tris gels (Thermo Fisher Scientific), Krypton (Thermo Fisher Scientific) staining and fluorescence laser scanning on a Bio-Rad FX Pro Plus Imager. .. 500 ng protein digest of each sample yielding similar total ion chromatograms (cf. in the SI) were injected and analyzed by nano-HPLC (Dionex Ultimate 3000) equipped with a C18, 5 μm, 100 Å, 5 × 0.3 mm, enrichment column and an Acclaim PepMap RSLC nanocolumn (C18, 2 μm, 100 Å, 500 × 0.075 mm) (all Thermo Fisher Scientific).

    Isolation:

    Article Title: Mechanisms of Yersinia YopO kinase substrate specificity
    Article Snippet: LC/MS analysis for in vitro phosphorylation Vacuum dried samples were reconstituted in 0.1% formic acid and analyzed using a nano-HPLC attached to an LTQ Orbitrap classic (Thermo Fisher Scientific). .. Ten of the most intense peptide ions in each survey scan with an ion intensity of > 2000 counts and a charge state ≥2 were isolated sequentially with a target value of 5000.

    Article Title: Yersinia effector protein (YopO)-mediated phosphorylation of host gelsolin causes calcium-independent activation leading to disruption of actin dynamics
    Article Snippet: Vacuum-dried samples were reconstituted in 0.1% formic acid and analyzed using a nano-HPLC coupled to an LTQ Orbitrap classic (Thermo Fisher Scientific). .. The 10 most intense peptide ions in each survey scan with an ion intensity of > 2000 counts and a charge state of ≥2 were isolated sequentially to a target value of 5000 and fragmented in the linear ion trap by collision-induced dissociation using a normalized collision energy of 35%.

    Purification:

    Article Title: Identification of potential mitochondrial CLPXP protease interactors and substrates suggests its central role in energy metabolism
    Article Snippet: Subsequently, samples were centrifuged at 1,000 g for 5 min at room temperature, acidified using 1% formic acid and purified using C18-ZipTips (Merck Millipore) according to the manufacturer’s protocol. .. Proteolytic digests were dissolved in 5% acetonitrile with 0.1% formic acid and loaded on reverse phase columns (trapping column: particle size 3 μm, C18, L = 20 mm; analytical column: particle size < 2 μm, C18, L = 50 cm; PepMap, Dionex/Thermo Fisher Scientific) using a nano-HPLC (Dionex – UltiMate 3000 RSLCnano, Thermo Fisher Scientific).

    Peptide Fractionation:

    Article Title: Proteome Analysis for Understanding Abiotic Stress (Salinity and Drought) Tolerance in Date Palm (Phoenix dactylifera L.)
    Article Snippet: The resulting peptides were analyzed by nano-HPLC (UltiMate 3000 HPLC System, LC Packings, Dionex, Idstein, Germany) coupled to an amaZon ETD MS ion trap spectrometer (Bruker Daltonics, Bremen, Germany) using nano-ESI spray. .. The liquid chromatography system was supplied with reversed-phase precolumn (LC Packings, Dionex) for sample desalting and a 15 cm PepMap 100 reversed-phase C18 column, 75 μ m inner diameter (LC Packings, Dionex), for peptide fractionation.

    Liquid Chromatography:

    Article Title: Relative Quantitative Proteomic Analysis of Brucella abortus Reveals Metabolic Adaptation to Multiple Environmental Stresses
    Article Snippet: .. Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) All experiments were performed on an LTQ Q-Exactive HF mass spectrometer (Thermo Scientific, USA) coupled online with a nano-HPLC (Ultimate 3000, Thermo Scientific) (Scheltema et al., ). .. The peptides were loaded onto a trap column (C18, 3 μm particles, 100 μm × 2 cm) and separated on EASY-Spray columns (C18, 1.9 μm particles, 15 μm × 12 cm) with trapping at a flow rate of 600 nL/min (Kentache et al., ).

    Article Title: MEIS homeodomain proteins facilitate PARP1/ARTD1-mediated eviction of histone H1
    Article Snippet: .. Liquid chromatography tandem mass spectrometry analyses The proteolytic digests were loaded using a nano-HPLC (Dionex RSLCnano) on reverse-phase columns (trapping column: Acclaim PepMap c18, particle size 2 µm, L = 20 mm; analytical column: Acclaim PepMap c18, particle size 2 µm, L = 25 cm; Thermo Fisher Scientific) and eluted in organic phase gradients (Buffer A: 95% H2 O, 5% DMSO, and 0.1% formic acid; Buffer B: 80% acetonitrile, 15% H2 O, 5% DMSO, and 0.1% formic acid). ..

    Article Title: Proteome Analysis for Understanding Abiotic Stress (Salinity and Drought) Tolerance in Date Palm (Phoenix dactylifera L.)
    Article Snippet: The resulting peptides were analyzed by nano-HPLC (UltiMate 3000 HPLC System, LC Packings, Dionex, Idstein, Germany) coupled to an amaZon ETD MS ion trap spectrometer (Bruker Daltonics, Bremen, Germany) using nano-ESI spray. .. The liquid chromatography system was supplied with reversed-phase precolumn (LC Packings, Dionex) for sample desalting and a 15 cm PepMap 100 reversed-phase C18 column, 75 μ m inner diameter (LC Packings, Dionex), for peptide fractionation.

    Software:

    Article Title: Buserelin alleviates chloride transport defect in human cystic fibrosis nasal epithelial cells
    Article Snippet: Gel alignment, spots detection and quantification were done using the PDQuest software (PDQuest Basic-8.0.1, BioRad, USA). .. Nano HPLC (Dionex RSLC UltiMate3000) was used, followed by MS (Scan range: 400–1500 m/z, fragmentation: CID, Energy of collision: 35%, Cycle for fragmentation (TopN): Top8).

    Article Title: Oligoribonuclease is a common downstream target of lithium-induced pAp accumulation in Escherichia coli and human cells
    Article Snippet: Bands corresponding to proteins binding specifically to pAp were localized by staining with Bio-Safe colloidal coomassie (Bio-Rad), cut out and digested in-gel by trypsin after reduction and alkylation according to standard protocols and analyzed by nano-HPLC (LC Packing) directly coupled to an ion trap mass spectrometer (ThermoFinnigan LCQ Deca XP) equipped with a nanoelectrospray source (LC-MS/MS). .. Database searches and evaluation of the results was done using the Bioworks 3.1 software.

    Article Title: Proteome Analysis for Understanding Abiotic Stress (Salinity and Drought) Tolerance in Date Palm (Phoenix dactylifera L.)
    Article Snippet: The resulting peptides were analyzed by nano-HPLC (UltiMate 3000 HPLC System, LC Packings, Dionex, Idstein, Germany) coupled to an amaZon ETD MS ion trap spectrometer (Bruker Daltonics, Bremen, Germany) using nano-ESI spray. .. The nano-HPLC system and the ion trap spectrometer were controlled using the Bruker Compass HyStar v3.2-SR2 software.

    Sample Prep:

    Article Title: Comparative proteomics of paired vocal fold and oral mucosa fibroblasts
    Article Snippet: For LC-MS/MS analysis 20 μg protein were subjected to filter aided sample preparation as described previously [ ] with minor modifications using 3 kD cut off ultracentrifugation filters (Millipore). .. 500 ng protein digest of each sample yielding similar total ion chromatograms (cf. in the SI) were injected and analyzed by nano-HPLC (Dionex Ultimate 3000) equipped with a C18, 5 μm, 100 Å, 5 × 0.3 mm, enrichment column and an Acclaim PepMap RSLC nanocolumn (C18, 2 μm, 100 Å, 500 × 0.075 mm) (all Thermo Fisher Scientific).

    In Vitro:

    Article Title: Mechanisms of Yersinia YopO kinase substrate specificity
    Article Snippet: .. LC/MS analysis for in vitro phosphorylation Vacuum dried samples were reconstituted in 0.1% formic acid and analyzed using a nano-HPLC attached to an LTQ Orbitrap classic (Thermo Fisher Scientific). ..

    Immunoprecipitation:

    Article Title: β3GnT8 Promotes Gastric Cancer Invasion by Regulating the Glycosylation of CD147
    Article Snippet: LC-MS/MS analysis Coomassie brilliant blue R-250 (Sigma) staining was performed to visually detect specific immunoprecipitated bands of β3GnT8 antibody. .. Digested peptides were analyzed by nano-HPLC (Ultimate 3000, Dionex) coupled to a linear quadrupole ion trap-Orbitrap (LTQ Orbitrap XL)mass spectrometer (Thermo Fisher) equipped with a nano-ESI source .

    Two-Dimensional Gel Electrophoresis:

    Article Title: Buserelin alleviates chloride transport defect in human cystic fibrosis nasal epithelial cells
    Article Snippet: Paragraph title: Two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) ... Nano HPLC (Dionex RSLC UltiMate3000) was used, followed by MS (Scan range: 400–1500 m/z, fragmentation: CID, Energy of collision: 35%, Cycle for fragmentation (TopN): Top8).

    Staining:

    Article Title: Buserelin alleviates chloride transport defect in human cystic fibrosis nasal epithelial cells
    Article Snippet: The 2-DE gels were stained with a G-250 Coomassie blue solution, scanned and digitalized using a GS-800 Calibrated Densitometer (Bio-Rad, USA). .. Nano HPLC (Dionex RSLC UltiMate3000) was used, followed by MS (Scan range: 400–1500 m/z, fragmentation: CID, Energy of collision: 35%, Cycle for fragmentation (TopN): Top8).

    Article Title: β3GnT8 Promotes Gastric Cancer Invasion by Regulating the Glycosylation of CD147
    Article Snippet: LC-MS/MS analysis Coomassie brilliant blue R-250 (Sigma) staining was performed to visually detect specific immunoprecipitated bands of β3GnT8 antibody. .. Digested peptides were analyzed by nano-HPLC (Ultimate 3000, Dionex) coupled to a linear quadrupole ion trap-Orbitrap (LTQ Orbitrap XL)mass spectrometer (Thermo Fisher) equipped with a nano-ESI source .

    Article Title: Oligoribonuclease is a common downstream target of lithium-induced pAp accumulation in Escherichia coli and human cells
    Article Snippet: .. Bands corresponding to proteins binding specifically to pAp were localized by staining with Bio-Safe colloidal coomassie (Bio-Rad), cut out and digested in-gel by trypsin after reduction and alkylation according to standard protocols and analyzed by nano-HPLC (LC Packing) directly coupled to an ion trap mass spectrometer (ThermoFinnigan LCQ Deca XP) equipped with a nanoelectrospray source (LC-MS/MS). .. Database searches and evaluation of the results was done using the Bioworks 3.1 software.

    Article Title: Comparative proteomics of paired vocal fold and oral mucosa fibroblasts
    Article Snippet: Protein patterns a were visualized by SDS-PAGE on 4–12% NuPAGE Bis-Tris gels (Thermo Fisher Scientific), Krypton (Thermo Fisher Scientific) staining and fluorescence laser scanning on a Bio-Rad FX Pro Plus Imager. .. 500 ng protein digest of each sample yielding similar total ion chromatograms (cf. in the SI) were injected and analyzed by nano-HPLC (Dionex Ultimate 3000) equipped with a C18, 5 μm, 100 Å, 5 × 0.3 mm, enrichment column and an Acclaim PepMap RSLC nanocolumn (C18, 2 μm, 100 Å, 500 × 0.075 mm) (all Thermo Fisher Scientific).

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  • 91
    Thermo Fisher high resolution nano hplc tandem mass spectrometry analysis
    Scheme of label-free quantitative proteomics. Synchronized animals at L4 stage were either fed E. Coli OP50 (uninfected controls) or P. aeruginosa PA14 (infected samples) for 4 hr, collected, and lysed. Total proteins were extracted and digested with trypsin. Peptides from control or infected samples were subject to <t>nano-HPLC</t> tandem MS analysis. Quantification is based on the comparison of peak intensity of same peptides in different samples.
    High Resolution Nano Hplc Tandem Mass Spectrometry Analysis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/high resolution nano hplc tandem mass spectrometry analysis/product/Thermo Fisher
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    high resolution nano hplc tandem mass spectrometry analysis - by Bioz Stars, 2020-04
    91/100 stars
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    91
    Thermo Fisher ultimate nano hplc system
    <t>Nano-HPLC/nano-ESI-LTQ-Orbitrap-MS/MS</t> analysis of cross-linked peptides derived from nidogen-1 G2 and G3 domain. The cross-linked product comprises amino acids 407–420 of the G2 domain (α-peptide, red) and 939–949 of the G3 domain (β-peptide, green), in which K-407 is connected to K-948/949.
    Ultimate Nano Hplc System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ultimate nano hplc system/product/Thermo Fisher
    Average 91 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    ultimate nano hplc system - by Bioz Stars, 2020-04
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    99
    Thermo Fisher nano hplc
    Experimental design of the quantitative proteomics of Brucella abortus under environmental stress. A label-free relative quantitative proteomics approach was utilized to investigate and compare the global proteomic changes of B. abortus in response to a variety of distinct stresses including a control condition, seven single-stress conditions, and a multi-stress condition. Protein samples were prepared and proteolytic-digested using trypsin enzymes. Peptides were analyzed on a Q-Exactive HF MS coupled online with a <t>nano-HPLC.</t> The identified proteins were quantified using a label-free approach and further functionally analyzed using the COG and KEGG databases.
    Nano Hplc, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 132 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nano hplc/product/Thermo Fisher
    Average 99 stars, based on 132 article reviews
    Price from $9.99 to $1999.99
    nano hplc - by Bioz Stars, 2020-04
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    95
    Thermo Fisher nano hplc system
    <t>Nano-HPLC/nano-ESI-LTQ-Orbitrap-MS/MS</t> analysis of cross-linked peptides derived from nidogen-1 G2 and G3 domain. The cross-linked product comprises amino acids 407–420 of the G2 domain (α-peptide, red) and 939–949 of the G3 domain (β-peptide, green), in which K-407 is connected to K-948/949.
    Nano Hplc System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nano hplc system/product/Thermo Fisher
    Average 95 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    nano hplc system - by Bioz Stars, 2020-04
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    Image Search Results


    Scheme of label-free quantitative proteomics. Synchronized animals at L4 stage were either fed E. Coli OP50 (uninfected controls) or P. aeruginosa PA14 (infected samples) for 4 hr, collected, and lysed. Total proteins were extracted and digested with trypsin. Peptides from control or infected samples were subject to nano-HPLC tandem MS analysis. Quantification is based on the comparison of peak intensity of same peptides in different samples.

    Journal: Scientific Reports

    Article Title: Neuronal GPCR OCTR-1 regulates innate immunity by controlling protein synthesis in Caenorhabditis elegans

    doi: 10.1038/srep36832

    Figure Lengend Snippet: Scheme of label-free quantitative proteomics. Synchronized animals at L4 stage were either fed E. Coli OP50 (uninfected controls) or P. aeruginosa PA14 (infected samples) for 4 hr, collected, and lysed. Total proteins were extracted and digested with trypsin. Peptides from control or infected samples were subject to nano-HPLC tandem MS analysis. Quantification is based on the comparison of peak intensity of same peptides in different samples.

    Article Snippet: High resolution nano-HPLC tandem mass spectrometry analysis The peptide samples were subjected to Thermo Scientific Orbitrap Fusion Tribrid with an Easy-nLC 1000 ultra-high pressure LC on a Thermo Scientific PepMap 100 C18 column (2 μm, 50 μm × 15 cm).

    Techniques: Infection, High Performance Liquid Chromatography, Mass Spectrometry

    Nano-HPLC/nano-ESI-LTQ-Orbitrap-MS/MS analysis of cross-linked peptides derived from nidogen-1 G2 and G3 domain. The cross-linked product comprises amino acids 407–420 of the G2 domain (α-peptide, red) and 939–949 of the G3 domain (β-peptide, green), in which K-407 is connected to K-948/949.

    Journal: PLoS ONE

    Article Title: Analysis of Nidogen-1/Laminin γ1 Interaction by Cross-Linking, Mass Spectrometry, and Computational Modeling Reveals Multiple Binding Modes

    doi: 10.1371/journal.pone.0112886

    Figure Lengend Snippet: Nano-HPLC/nano-ESI-LTQ-Orbitrap-MS/MS analysis of cross-linked peptides derived from nidogen-1 G2 and G3 domain. The cross-linked product comprises amino acids 407–420 of the G2 domain (α-peptide, red) and 939–949 of the G3 domain (β-peptide, green), in which K-407 is connected to K-948/949.

    Article Snippet: Nano-HPLC/Nano-ESI-LTQ-Orbitrap-MS/MS Fractionation of peptide mixtures was carried out on an Ultimate nano-HPLC system (Thermo Fisher Scientific) using reversed phase C18 columns (precolumn: Acclaim PepMap, 300 µm • 5 mm, 5 µm, 100 Å, separation column: Acclaim PepMap, 75 µm • 250 mm, 3 µm, 100 Å, Thermo Fisher Scientific).

    Techniques: High Performance Liquid Chromatography, Mass Spectrometry, Derivative Assay

    Experimental design of the quantitative proteomics of Brucella abortus under environmental stress. A label-free relative quantitative proteomics approach was utilized to investigate and compare the global proteomic changes of B. abortus in response to a variety of distinct stresses including a control condition, seven single-stress conditions, and a multi-stress condition. Protein samples were prepared and proteolytic-digested using trypsin enzymes. Peptides were analyzed on a Q-Exactive HF MS coupled online with a nano-HPLC. The identified proteins were quantified using a label-free approach and further functionally analyzed using the COG and KEGG databases.

    Journal: Frontiers in Microbiology

    Article Title: Relative Quantitative Proteomic Analysis of Brucella abortus Reveals Metabolic Adaptation to Multiple Environmental Stresses

    doi: 10.3389/fmicb.2017.02347

    Figure Lengend Snippet: Experimental design of the quantitative proteomics of Brucella abortus under environmental stress. A label-free relative quantitative proteomics approach was utilized to investigate and compare the global proteomic changes of B. abortus in response to a variety of distinct stresses including a control condition, seven single-stress conditions, and a multi-stress condition. Protein samples were prepared and proteolytic-digested using trypsin enzymes. Peptides were analyzed on a Q-Exactive HF MS coupled online with a nano-HPLC. The identified proteins were quantified using a label-free approach and further functionally analyzed using the COG and KEGG databases.

    Article Snippet: Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) All experiments were performed on an LTQ Q-Exactive HF mass spectrometer (Thermo Scientific, USA) coupled online with a nano-HPLC (Ultimate 3000, Thermo Scientific) (Scheltema et al., ).

    Techniques: Mass Spectrometry, High Performance Liquid Chromatography

    Nano-HPLC/nano-ESI-LTQ-Orbitrap-MS/MS analysis of cross-linked peptides derived from nidogen-1 G2 and G3 domain. The cross-linked product comprises amino acids 407–420 of the G2 domain (α-peptide, red) and 939–949 of the G3 domain (β-peptide, green), in which K-407 is connected to K-948/949.

    Journal: PLoS ONE

    Article Title: Analysis of Nidogen-1/Laminin γ1 Interaction by Cross-Linking, Mass Spectrometry, and Computational Modeling Reveals Multiple Binding Modes

    doi: 10.1371/journal.pone.0112886

    Figure Lengend Snippet: Nano-HPLC/nano-ESI-LTQ-Orbitrap-MS/MS analysis of cross-linked peptides derived from nidogen-1 G2 and G3 domain. The cross-linked product comprises amino acids 407–420 of the G2 domain (α-peptide, red) and 939–949 of the G3 domain (β-peptide, green), in which K-407 is connected to K-948/949.

    Article Snippet: The nano-HPLC system was directly coupled to the nano-ESI source (Proxeon) of an LTQ-Orbitrap XL hybrid mass spectrometer (Thermo Fisher Scientific).

    Techniques: High Performance Liquid Chromatography, Mass Spectrometry, Derivative Assay