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Thermo Fisher nanohplc
LC-ESI-MS/MS analysis of fengycins from Bacillus amyloliquefaciens MEP 2 18. Elution profile of fengycins (A) , obtained from a HPLC-purified peak after separation of CLP from B. amyloliquefaciens MEP 2 18, using <t>nanoHPLC</t> technology (Thermo Scientific, EASY-nLC 1000) equipped with a reverse-phase C18 column (Easy-Spray ColumnPepMap RSLC, P/N ES803, 2 μm particle size, 75 μm × 500 mm, Thermo Scientific). ESI-MS/MS spectra of protonated fengycin ions m/z 1463.8 (B) obtained from peak 1, the spectrum of precursor ion of m/z 1463.8 (C) ; m/z 1477.8 (D) obtained from peak 2, the spectrum of precursor ion of m/z 1477.8 (E) ; and ions m/z 1477.8, 1491.8, and 1505.8 (F) obtained from peak 3, the spectrum of precursor ion of m/z 1477.8 (G) , 1491.8 (H) , and 1505.8 (I) were acquired in a mass spectrometer (Thermo Scientific, Q-Exactive) with a high collision dissociation (HCD) cell and an Orbitrap analyzer.
Nanohplc, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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89/100 stars

Images

1) Product Images from "Fengycins From Bacillus amyloliquefaciens MEP218 Exhibit Antibacterial Activity by Producing Alterations on the Cell Surface of the Pathogens Xanthomonas axonopodis pv. vesicatoria and Pseudomonas aeruginosa PA01"

Article Title: Fengycins From Bacillus amyloliquefaciens MEP218 Exhibit Antibacterial Activity by Producing Alterations on the Cell Surface of the Pathogens Xanthomonas axonopodis pv. vesicatoria and Pseudomonas aeruginosa PA01

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2019.03107

LC-ESI-MS/MS analysis of fengycins from Bacillus amyloliquefaciens MEP 2 18. Elution profile of fengycins (A) , obtained from a HPLC-purified peak after separation of CLP from B. amyloliquefaciens MEP 2 18, using nanoHPLC technology (Thermo Scientific, EASY-nLC 1000) equipped with a reverse-phase C18 column (Easy-Spray ColumnPepMap RSLC, P/N ES803, 2 μm particle size, 75 μm × 500 mm, Thermo Scientific). ESI-MS/MS spectra of protonated fengycin ions m/z 1463.8 (B) obtained from peak 1, the spectrum of precursor ion of m/z 1463.8 (C) ; m/z 1477.8 (D) obtained from peak 2, the spectrum of precursor ion of m/z 1477.8 (E) ; and ions m/z 1477.8, 1491.8, and 1505.8 (F) obtained from peak 3, the spectrum of precursor ion of m/z 1477.8 (G) , 1491.8 (H) , and 1505.8 (I) were acquired in a mass spectrometer (Thermo Scientific, Q-Exactive) with a high collision dissociation (HCD) cell and an Orbitrap analyzer.
Figure Legend Snippet: LC-ESI-MS/MS analysis of fengycins from Bacillus amyloliquefaciens MEP 2 18. Elution profile of fengycins (A) , obtained from a HPLC-purified peak after separation of CLP from B. amyloliquefaciens MEP 2 18, using nanoHPLC technology (Thermo Scientific, EASY-nLC 1000) equipped with a reverse-phase C18 column (Easy-Spray ColumnPepMap RSLC, P/N ES803, 2 μm particle size, 75 μm × 500 mm, Thermo Scientific). ESI-MS/MS spectra of protonated fengycin ions m/z 1463.8 (B) obtained from peak 1, the spectrum of precursor ion of m/z 1463.8 (C) ; m/z 1477.8 (D) obtained from peak 2, the spectrum of precursor ion of m/z 1477.8 (E) ; and ions m/z 1477.8, 1491.8, and 1505.8 (F) obtained from peak 3, the spectrum of precursor ion of m/z 1477.8 (G) , 1491.8 (H) , and 1505.8 (I) were acquired in a mass spectrometer (Thermo Scientific, Q-Exactive) with a high collision dissociation (HCD) cell and an Orbitrap analyzer.

Techniques Used: Tandem Mass Spectroscopy, High Performance Liquid Chromatography, Purification, Mass Spectrometry

2) Product Images from "Proteome profiling of s-nitrosylated synaptosomal proteins by isobaric mass tags"

Article Title: Proteome profiling of s-nitrosylated synaptosomal proteins by isobaric mass tags

Journal: Journal of Neuroscience Methods

doi: 10.1016/j.jneumeth.2017.08.005

The isobaric tag labeling procedure. (A) Schematic image of the isobaric tag labeling procedure. After synaptosome isolation, SNO proteins are labeled by (i) blocking of free thiols using MMTS; (ii) reduction of cysteine S -nitrosylation with ascorbate, producing new free thiols on previous SNO sites; and (iii) isobaric tag labeling of the new thiols using iodoTMT. iodoTMT-labeled proteins are subjected to in-solution protein digestion. Derived peptides are enriched using affinity-capture on an anti-TMT resin. Eluted peptides are identified by nanoHPLC–MS/MS and quantified using the TMT sixplex. (B) A representative MS/MS spectrum of the iodoTMT-labeled peptide SICDTSNFSDYIR derived from ubiquitin-like modifier-activating enzyme 1. Peaks of the 6 reporter ions are found in the enlarged section of the lower mass range.
Figure Legend Snippet: The isobaric tag labeling procedure. (A) Schematic image of the isobaric tag labeling procedure. After synaptosome isolation, SNO proteins are labeled by (i) blocking of free thiols using MMTS; (ii) reduction of cysteine S -nitrosylation with ascorbate, producing new free thiols on previous SNO sites; and (iii) isobaric tag labeling of the new thiols using iodoTMT. iodoTMT-labeled proteins are subjected to in-solution protein digestion. Derived peptides are enriched using affinity-capture on an anti-TMT resin. Eluted peptides are identified by nanoHPLC–MS/MS and quantified using the TMT sixplex. (B) A representative MS/MS spectrum of the iodoTMT-labeled peptide SICDTSNFSDYIR derived from ubiquitin-like modifier-activating enzyme 1. Peaks of the 6 reporter ions are found in the enlarged section of the lower mass range.

Techniques Used: Labeling, Isolation, Blocking Assay, Derivative Assay, Mass Spectrometry

3) Product Images from "Quantification of Cardiovascular Disease Biomarkers in Human Platelets by Targeted Mass Spectrometry"

Article Title: Quantification of Cardiovascular Disease Biomarkers in Human Platelets by Targeted Mass Spectrometry

Journal: Proteomes

doi: 10.3390/proteomes5040031

Quantification of platelet proteins by an internal standard curve. ( A ) Proteomics workflow. The prepared peptide samples were separated by nanoHPLC and measured subsequently with nanoESI-MS parallel reaction monitoring (PRM). Peptides of interest were isolated sequentially in a quadrupole, then fragmented, and finally peptide fragments detected in an orbitrap mass analyzer; ( B ) Distribution of endogenous concentration of quantified proteins. In total, 133 peptides corresponding to 99 proteins in human platelets were quantified; ( C ) Quantification of Integrin alpha-2 (ITA2) via an internal calibration curve and least squares linear regression. The dots represent the measured areas of the quantifier transition of the stable isotope-labeled (black) and endogenous peptides (red) plotted against the known or determined peptide concentration. Plotted in grey is the straight line fitted to the measured area under the curve of the quantifier transition of stable isotope-labeled peptide using least squares linear regression.
Figure Legend Snippet: Quantification of platelet proteins by an internal standard curve. ( A ) Proteomics workflow. The prepared peptide samples were separated by nanoHPLC and measured subsequently with nanoESI-MS parallel reaction monitoring (PRM). Peptides of interest were isolated sequentially in a quadrupole, then fragmented, and finally peptide fragments detected in an orbitrap mass analyzer; ( B ) Distribution of endogenous concentration of quantified proteins. In total, 133 peptides corresponding to 99 proteins in human platelets were quantified; ( C ) Quantification of Integrin alpha-2 (ITA2) via an internal calibration curve and least squares linear regression. The dots represent the measured areas of the quantifier transition of the stable isotope-labeled (black) and endogenous peptides (red) plotted against the known or determined peptide concentration. Plotted in grey is the straight line fitted to the measured area under the curve of the quantifier transition of stable isotope-labeled peptide using least squares linear regression.

Techniques Used: Mass Spectrometry, Isolation, Concentration Assay, Labeling

4) Product Images from "High-Resolution Mass Spectrometry Driven Discovery of Peptidic Danger Signals in Insect Immunity"

Article Title: High-Resolution Mass Spectrometry Driven Discovery of Peptidic Danger Signals in Insect Immunity

Journal: PLoS ONE

doi: 10.1371/journal.pone.0080406

Workflow for the identification of bioactive peptides. On the one hand the thermolysin-digested hemolymph sample was analyzed directly by nanoHPLC-FTMS. On the other hand the thermolysin-digested sample was pre-fractionated using a HPLC-system and the collected samples were tested for bioactivity for a more detailed analysis of immuno-relevant peptides.
Figure Legend Snippet: Workflow for the identification of bioactive peptides. On the one hand the thermolysin-digested hemolymph sample was analyzed directly by nanoHPLC-FTMS. On the other hand the thermolysin-digested sample was pre-fractionated using a HPLC-system and the collected samples were tested for bioactivity for a more detailed analysis of immuno-relevant peptides.

Techniques Used: High Performance Liquid Chromatography

Related Articles

Mass Spectrometry:

Article Title: CDC20B is required for deuterosome-mediated centriole production in multiciliated cells
Article Snippet: .. NanoHPLC was coupled via a nanoelectrospray ionization source to the Hybrid Quadrupole-Orbitrap High Resolution Mass Spectrometer (ThermoFisher Scientific). .. MS spectra were acquired at a resolution of 70,000 (200 m /z ) in a mass range of 300–2000 m /z with an AGC target 3e6 value of and a maximum injection time of 100 ms.

Article Title: Proteome profiling of s-nitrosylated synaptosomal proteins by isobaric mass tags
Article Snippet: .. The nanoHPLC was coupled online to an LTQ Orbitrap Velos mass spectrometer (Thermo Fisher Scientific, Bremen, Germany). .. Peptide ions between 330 and 1700 m /z were scanned in the Orbitrap detector with a resolution of 30,000 (maximum fill time 400 ms, AGC target 106).

Article Title: How recombinant swollenin from Kluyveromyces lactis affects cellulosic substrates and accelerates their hydrolysis
Article Snippet: .. Peptide analysis was carried out using a nanoHPLC (Dionex, Germering, Germany) coupled to an ESI-QUAD-TOF-2 mass spectrometer (Waters Micromass, Eschborn, Germany) as previously described [ ]. .. The Mascot algorithm (Matrix Science, London, UK) was used to correlate the mass spectrometry data with amino acid sequences in the Swissprot database.

Article Title: TOM1L1 drives membrane delivery of MT1-MMP to promote ERBB2-induced breast cancer cell invasion
Article Snippet: .. Briefly, samples were analysed using nanoHPLC (Ultimate 3000, Dionex)/nanoelectrospray ionization on an orbitrap mass spectrometer (LTQ-Orbitrap XL, Thermo Fischer scientific). .. Sample desalting and pre-concentration were carried out online using a Pepmapper column (0.3 × 10 mm, Dionex).

Article Title: High-Resolution Mass Spectrometry Driven Discovery of Peptidic Danger Signals in Insect Immunity
Article Snippet: .. In addition, coupling of nanoHPLC with a linear ion trap / Fourier transform orbital trapping (IT-FTOT MS) mass spectrometer (LTQ Orbitrap Discovery, Thermo Scientific GmbH , Bremen, Germany) equipped with a nanospray ion source, was used. .. This mass spectrometer allowed, in addition to CID-fragmentation, the detection of low mass fragments by higher-energy collisional dissociation (HCD), providing additional information for peptide sequencing.

Article Title: Quantification of Cardiovascular Disease Biomarkers in Human Platelets by Targeted Mass Spectrometry
Article Snippet: .. The nanoHPLC was coupled to a Q Exactive Hybrid Quadrupole-Orbitrap Plus mass spectrometer (Thermo Scientific) and eluting peptides were ionized directly via a silica emitter (FS360-20-10-D-20, New Objective, Ringoes, NJ, USA) of the nanoESI source. .. Peptides of interest were analyzed using a scheduled parallel reaction monitoring method, which was performed with a resolution of 17,500, a maximum injection time of 50 ms, and an automatic gain control (AGC) value of 3 × 106 .

Article Title: TRIAD3/RNF216 E3 ligase specifically synthesises K63-linked ubiquitin chains and is inactivated by mutations associated with Gordon Holmes syndrome
Article Snippet: .. The nanoHPLC was coupled online to an LTQ Orbitrap Velos mass spectrometer (Thermo Fisher Scientific). .. Peptide ions between 330 and 1600 m/z were scanned in the Orbitrap detector with a resolution of 30,000 (maximum fill time 400 ms, AGC target 106).

Purification:

Article Title: Fengycins From Bacillus amyloliquefaciens MEP218 Exhibit Antibacterial Activity by Producing Alterations on the Cell Surface of the Pathogens Xanthomonas axonopodis pv. vesicatoria and Pseudomonas aeruginosa PA01
Article Snippet: .. Fengycins were purified with ZipTips C18 and analyzed by nanoHPLC (Thermo Scientific, EASY-nLC 1000) equipped with a reverse-phase C18 column (Easy-Spray ColumnPepMap RSLC, P/N ES803, 2 μm particle size, 75 μm × 500 mm, Thermo Scientific) at 35°C. .. A two component solvent system was used: solvent A is water (HPLC grade) acidified with 0.1% formic acid and solvent B is acetonitrile with 0.1% formic acid.

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    Thermo Fisher nanohplc
    LC-ESI-MS/MS analysis of fengycins from Bacillus amyloliquefaciens MEP 2 18. Elution profile of fengycins (A) , obtained from a HPLC-purified peak after separation of CLP from B. amyloliquefaciens MEP 2 18, using <t>nanoHPLC</t> technology (Thermo Scientific, EASY-nLC 1000) equipped with a reverse-phase C18 column (Easy-Spray ColumnPepMap RSLC, P/N ES803, 2 μm particle size, 75 μm × 500 mm, Thermo Scientific). ESI-MS/MS spectra of protonated fengycin ions m/z 1463.8 (B) obtained from peak 1, the spectrum of precursor ion of m/z 1463.8 (C) ; m/z 1477.8 (D) obtained from peak 2, the spectrum of precursor ion of m/z 1477.8 (E) ; and ions m/z 1477.8, 1491.8, and 1505.8 (F) obtained from peak 3, the spectrum of precursor ion of m/z 1477.8 (G) , 1491.8 (H) , and 1505.8 (I) were acquired in a mass spectrometer (Thermo Scientific, Q-Exactive) with a high collision dissociation (HCD) cell and an Orbitrap analyzer.
    Nanohplc, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nanohplc/product/Thermo Fisher
    Average 89 stars, based on 44 article reviews
    Price from $9.99 to $1999.99
    nanohplc - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

    91
    Thermo Fisher nanohplc maldi ms ms
    Pull-down experiment of serum pre-incubated dPGS-Au50 or dPGOH-Au50 with purified membrane fractions. Notes: The dPGS-Au50 ( A ) and dPGOH-Au50 ( B ) NPs were incubated with RPMI-1640 medium serum containing 10% FBS (CCM), with isolated MPs or were first pre-incubated with RPMI-1640 medium containing 10% FBS and then incubated with MPs (CCM + MPs). Proteins were eluted from NP surfaces using Laemmli buffer and analyzed by SDS-PAGE. Eluted proteins were visualized by Coomassie and the identified by <t>nanoHPLC</t> coupled with <t>MALDI-TOF/TOF.</t> Abbreviations: NP, nanoparticle; FBS, fetal bovine serum; MPs, membrane proteins; CCM, complete cell culture medium; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; nanoHPLC, nano high performance liquid chromatography; MALDI, matrix-assisted laser desorption ionization; TOF, time of flight.
    Nanohplc Maldi Ms Ms, 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/nanohplc maldi ms ms/product/Thermo Fisher
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nanohplc maldi ms ms - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    Image Search Results


    LC-ESI-MS/MS analysis of fengycins from Bacillus amyloliquefaciens MEP 2 18. Elution profile of fengycins (A) , obtained from a HPLC-purified peak after separation of CLP from B. amyloliquefaciens MEP 2 18, using nanoHPLC technology (Thermo Scientific, EASY-nLC 1000) equipped with a reverse-phase C18 column (Easy-Spray ColumnPepMap RSLC, P/N ES803, 2 μm particle size, 75 μm × 500 mm, Thermo Scientific). ESI-MS/MS spectra of protonated fengycin ions m/z 1463.8 (B) obtained from peak 1, the spectrum of precursor ion of m/z 1463.8 (C) ; m/z 1477.8 (D) obtained from peak 2, the spectrum of precursor ion of m/z 1477.8 (E) ; and ions m/z 1477.8, 1491.8, and 1505.8 (F) obtained from peak 3, the spectrum of precursor ion of m/z 1477.8 (G) , 1491.8 (H) , and 1505.8 (I) were acquired in a mass spectrometer (Thermo Scientific, Q-Exactive) with a high collision dissociation (HCD) cell and an Orbitrap analyzer.

    Journal: Frontiers in Microbiology

    Article Title: Fengycins From Bacillus amyloliquefaciens MEP218 Exhibit Antibacterial Activity by Producing Alterations on the Cell Surface of the Pathogens Xanthomonas axonopodis pv. vesicatoria and Pseudomonas aeruginosa PA01

    doi: 10.3389/fmicb.2019.03107

    Figure Lengend Snippet: LC-ESI-MS/MS analysis of fengycins from Bacillus amyloliquefaciens MEP 2 18. Elution profile of fengycins (A) , obtained from a HPLC-purified peak after separation of CLP from B. amyloliquefaciens MEP 2 18, using nanoHPLC technology (Thermo Scientific, EASY-nLC 1000) equipped with a reverse-phase C18 column (Easy-Spray ColumnPepMap RSLC, P/N ES803, 2 μm particle size, 75 μm × 500 mm, Thermo Scientific). ESI-MS/MS spectra of protonated fengycin ions m/z 1463.8 (B) obtained from peak 1, the spectrum of precursor ion of m/z 1463.8 (C) ; m/z 1477.8 (D) obtained from peak 2, the spectrum of precursor ion of m/z 1477.8 (E) ; and ions m/z 1477.8, 1491.8, and 1505.8 (F) obtained from peak 3, the spectrum of precursor ion of m/z 1477.8 (G) , 1491.8 (H) , and 1505.8 (I) were acquired in a mass spectrometer (Thermo Scientific, Q-Exactive) with a high collision dissociation (HCD) cell and an Orbitrap analyzer.

    Article Snippet: Fengycins were purified with ZipTips C18 and analyzed by nanoHPLC (Thermo Scientific, EASY-nLC 1000) equipped with a reverse-phase C18 column (Easy-Spray ColumnPepMap RSLC, P/N ES803, 2 μm particle size, 75 μm × 500 mm, Thermo Scientific) at 35°C.

    Techniques: Tandem Mass Spectroscopy, High Performance Liquid Chromatography, Purification, Mass Spectrometry

    The isobaric tag labeling procedure. (A) Schematic image of the isobaric tag labeling procedure. After synaptosome isolation, SNO proteins are labeled by (i) blocking of free thiols using MMTS; (ii) reduction of cysteine S -nitrosylation with ascorbate, producing new free thiols on previous SNO sites; and (iii) isobaric tag labeling of the new thiols using iodoTMT. iodoTMT-labeled proteins are subjected to in-solution protein digestion. Derived peptides are enriched using affinity-capture on an anti-TMT resin. Eluted peptides are identified by nanoHPLC–MS/MS and quantified using the TMT sixplex. (B) A representative MS/MS spectrum of the iodoTMT-labeled peptide SICDTSNFSDYIR derived from ubiquitin-like modifier-activating enzyme 1. Peaks of the 6 reporter ions are found in the enlarged section of the lower mass range.

    Journal: Journal of Neuroscience Methods

    Article Title: Proteome profiling of s-nitrosylated synaptosomal proteins by isobaric mass tags

    doi: 10.1016/j.jneumeth.2017.08.005

    Figure Lengend Snippet: The isobaric tag labeling procedure. (A) Schematic image of the isobaric tag labeling procedure. After synaptosome isolation, SNO proteins are labeled by (i) blocking of free thiols using MMTS; (ii) reduction of cysteine S -nitrosylation with ascorbate, producing new free thiols on previous SNO sites; and (iii) isobaric tag labeling of the new thiols using iodoTMT. iodoTMT-labeled proteins are subjected to in-solution protein digestion. Derived peptides are enriched using affinity-capture on an anti-TMT resin. Eluted peptides are identified by nanoHPLC–MS/MS and quantified using the TMT sixplex. (B) A representative MS/MS spectrum of the iodoTMT-labeled peptide SICDTSNFSDYIR derived from ubiquitin-like modifier-activating enzyme 1. Peaks of the 6 reporter ions are found in the enlarged section of the lower mass range.

    Article Snippet: The nanoHPLC was coupled online to an LTQ Orbitrap Velos mass spectrometer (Thermo Fisher Scientific, Bremen, Germany).

    Techniques: Labeling, Isolation, Blocking Assay, Derivative Assay, Mass Spectrometry

    Quantification of platelet proteins by an internal standard curve. ( A ) Proteomics workflow. The prepared peptide samples were separated by nanoHPLC and measured subsequently with nanoESI-MS parallel reaction monitoring (PRM). Peptides of interest were isolated sequentially in a quadrupole, then fragmented, and finally peptide fragments detected in an orbitrap mass analyzer; ( B ) Distribution of endogenous concentration of quantified proteins. In total, 133 peptides corresponding to 99 proteins in human platelets were quantified; ( C ) Quantification of Integrin alpha-2 (ITA2) via an internal calibration curve and least squares linear regression. The dots represent the measured areas of the quantifier transition of the stable isotope-labeled (black) and endogenous peptides (red) plotted against the known or determined peptide concentration. Plotted in grey is the straight line fitted to the measured area under the curve of the quantifier transition of stable isotope-labeled peptide using least squares linear regression.

    Journal: Proteomes

    Article Title: Quantification of Cardiovascular Disease Biomarkers in Human Platelets by Targeted Mass Spectrometry

    doi: 10.3390/proteomes5040031

    Figure Lengend Snippet: Quantification of platelet proteins by an internal standard curve. ( A ) Proteomics workflow. The prepared peptide samples were separated by nanoHPLC and measured subsequently with nanoESI-MS parallel reaction monitoring (PRM). Peptides of interest were isolated sequentially in a quadrupole, then fragmented, and finally peptide fragments detected in an orbitrap mass analyzer; ( B ) Distribution of endogenous concentration of quantified proteins. In total, 133 peptides corresponding to 99 proteins in human platelets were quantified; ( C ) Quantification of Integrin alpha-2 (ITA2) via an internal calibration curve and least squares linear regression. The dots represent the measured areas of the quantifier transition of the stable isotope-labeled (black) and endogenous peptides (red) plotted against the known or determined peptide concentration. Plotted in grey is the straight line fitted to the measured area under the curve of the quantifier transition of stable isotope-labeled peptide using least squares linear regression.

    Article Snippet: The nanoHPLC was coupled to a Q Exactive Hybrid Quadrupole-Orbitrap Plus mass spectrometer (Thermo Scientific) and eluting peptides were ionized directly via a silica emitter (FS360-20-10-D-20, New Objective, Ringoes, NJ, USA) of the nanoESI source.

    Techniques: Mass Spectrometry, Isolation, Concentration Assay, Labeling

    Pull-down experiment of serum pre-incubated dPGS-Au50 or dPGOH-Au50 with purified membrane fractions. Notes: The dPGS-Au50 ( A ) and dPGOH-Au50 ( B ) NPs were incubated with RPMI-1640 medium serum containing 10% FBS (CCM), with isolated MPs or were first pre-incubated with RPMI-1640 medium containing 10% FBS and then incubated with MPs (CCM + MPs). Proteins were eluted from NP surfaces using Laemmli buffer and analyzed by SDS-PAGE. Eluted proteins were visualized by Coomassie and the identified by nanoHPLC coupled with MALDI-TOF/TOF. Abbreviations: NP, nanoparticle; FBS, fetal bovine serum; MPs, membrane proteins; CCM, complete cell culture medium; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; nanoHPLC, nano high performance liquid chromatography; MALDI, matrix-assisted laser desorption ionization; TOF, time of flight.

    Journal: International Journal of Nanomedicine

    Article Title: The influence of surface charge on serum protein interaction and cellular uptake: studies with dendritic polyglycerols and dendritic polyglycerol-coated gold nanoparticles

    doi: 10.2147/IJN.S124295

    Figure Lengend Snippet: Pull-down experiment of serum pre-incubated dPGS-Au50 or dPGOH-Au50 with purified membrane fractions. Notes: The dPGS-Au50 ( A ) and dPGOH-Au50 ( B ) NPs were incubated with RPMI-1640 medium serum containing 10% FBS (CCM), with isolated MPs or were first pre-incubated with RPMI-1640 medium containing 10% FBS and then incubated with MPs (CCM + MPs). Proteins were eluted from NP surfaces using Laemmli buffer and analyzed by SDS-PAGE. Eluted proteins were visualized by Coomassie and the identified by nanoHPLC coupled with MALDI-TOF/TOF. Abbreviations: NP, nanoparticle; FBS, fetal bovine serum; MPs, membrane proteins; CCM, complete cell culture medium; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; nanoHPLC, nano high performance liquid chromatography; MALDI, matrix-assisted laser desorption ionization; TOF, time of flight.

    Article Snippet: Analysis of pulled down proteins through nanoHPLC-MALDI-MS/MS For the chromatographic separation of the samples, a reversed-phase HPLC UltiMate3000 (Thermo Scientific) with gradient elution was used.

    Techniques: Incubation, Purification, Isolation, SDS Page, Cell Culture, Polyacrylamide Gel Electrophoresis, High Performance Liquid Chromatography