ltq-orbitrap mass spectrometer Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96
    Thermo Fisher maldi ltq orbitrap mass spectrometer
    Workflow for MS imaging of Pseudonocardia on the ant propleural plate. a) Photograph and b) cartoon of Pseudonocardia on the ant exoskeleton. c) Grooves were cut into glass slides and double-sided tape was applied to the back of the slide. The ant thoraxes were removed and positioned into the groove of the slide with the propleural plate facing outward and even with the top of the slide. An additional thin strip of double-sided tape was applied below the propleural plate to stabilize and secure the thorax into place. d) Matrix was applied to the slide using an automatic sprayer. e) A laser was fired at the sample to ionize compounds of interest and introduce them into the mass spectrometer. f) An array of mass spectra was acquired using a <t>MALDI-LTQ</t> <t>Orbitrap</t> and g) compiled into MS images. Photo courtesy of Alex Wild.
    Maldi Ltq Orbitrap Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/maldi ltq orbitrap mass spectrometer/product/Thermo Fisher
    Average 96 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    maldi ltq orbitrap mass spectrometer - by Bioz Stars, 2020-07
    96/100 stars
      Buy from Supplier

    95
    Thermo Fisher ltq orbitrap mass spectrometer
    Human Parkin Ser 65 is a substrate of human PINK1 upon CCCP stimulation. ( a ) Confirmation by mass spectrometry that Ser 65 of human Parkin is phosphorylated by CCCP-induced activation of human wild-type PINK1-FLAG. Flp-In T-Rex HEK293 cells expressing FLAG-empty, wild-type PINK1-FLAG, and kinase-inactive PINK1-FLAG (D384A) were co-transfected with HA-Parkin, induced with doxycycline and stimulated with 10 μM of CCCP for 3 h. Whole-cell extracts were obtained following lysis with 1% Triton and approximately 30 mg of whole-cell extract were subjected to immunoprecipitation with anti-HA-agarose and run on 10% SDS-PAGE and stained with colloidal Coomassie blue. Coomassie-stained bands migrating with the expected molecular mass of HA-Parkin were excised from the gel, digested with trypsin, and subjected to high performance liquid chromatography with tandem mass spectrometry (LC-MS-MS) on an <t>LTQ-Orbitrap</t> mass spectrometer. Extracted ion chromatogram analysis of Ser 131 and Ser 65 phosphopeptide (3 + R.NDWTVQNCDLDQQ S IVHIVQRPWR.K+P). The total signal intensity of the phosphopeptide is plotted on the y -axis and retention time is plotted on the x -axis. The m / z value corresponding to the Ser 131 phosphopeptide was detected in all conditions whilst that of the Ser 65 phosphopeptide was only detected in samples from wild-type PINK1-FLAG-expressing cells following CCCP treatment. ( b ) Characterization of Parkin phospho-Ser 65 antibody. Flp-In T-Rex HEK293 cells expressing FLAG-empty, wild-type PINK1-FLAG, and kinase-inactive PINK1-FLAG were co-transfected with untagged wild-type (WT) or Ser 65 Ala (S65A) mutant Parkin, induced with doxycycline and stimulated with 10 μM of CCCP for 3 h. 0.25 mg of 1% Triton whole-cell lysate were subjected to immunoprecipitation with anti-Parkin antibody (S966C) covalently coupled to protein G Sepharose and then immunoblotted with anti-phospho-Ser 65 antibody in the presence of dephosphorylated peptide. Ten per cent of the immunoprecipitate (IP) was immunoblotted with total anti-Parkin antibody. Twenty five micrograms of whole cell lysate was immunoblotted with total PINK1 antibody.
    Ltq Orbitrap Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 9541 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ltq orbitrap mass spectrometer/product/Thermo Fisher
    Average 95 stars, based on 9541 article reviews
    Price from $9.99 to $1999.99
    ltq orbitrap mass spectrometer - by Bioz Stars, 2020-07
    95/100 stars
      Buy from Supplier

    89
    Thermo Fisher maldi ltq orbitrap xl mass spectrometer
    Full scan spectra of protein mixtures: <t>MALDI-TOF</t> spectrum of histone mixture, 10 mg/mL (a), HRAM <t>MAI-Orbitrap</t> spectrum of histone mixture, 1 mg/mL (b) and HRAM MAI-Orbitrap spectrum of rat brain protein extract.
    Maldi Ltq Orbitrap Xl Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 134 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/maldi ltq orbitrap xl mass spectrometer/product/Thermo Fisher
    Average 89 stars, based on 134 article reviews
    Price from $9.99 to $1999.99
    maldi ltq orbitrap xl mass spectrometer - by Bioz Stars, 2020-07
    89/100 stars
      Buy from Supplier

    90
    Thermo Fisher ltq orbitrap discovery mass spectrometer
    Partial sequence (162–174) is present in unprocessed ABCA3 and missing in processed protein form. A : Intensity profiles of an m/z range corresponding to the monoisotopic peak of the doubly-charged ion at m/z 819.885 ( m/z window ±5 ppm) for 190 kDa and 170 kDa gel bands acquired in <t>Orbitrap.</t> B : Identification of the ABCA3 tryptic peptide comprising amino acid residues 162–174 of the precursor protein and containing methionine sulfoxide at position 164. MS/MS spectra after CID fragmentation measured in the <t>LTQ</t> ion trap are shown. Major b and y ions are labelled on the spectra and all fragment ions obtained are marked on the identified sequence. The ion at m/z 811 corresponds to the loss of water from the parent ion.
    Ltq Orbitrap Discovery Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 467 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ltq orbitrap discovery mass spectrometer/product/Thermo Fisher
    Average 90 stars, based on 467 article reviews
    Price from $9.99 to $1999.99
    ltq orbitrap discovery mass spectrometer - by Bioz Stars, 2020-07
    90/100 stars
      Buy from Supplier

    85
    Thermo Fisher 1100 ltq orbitrap mass spectrometer
    Partial sequence (162–174) is present in unprocessed ABCA3 and missing in processed protein form. A : Intensity profiles of an m/z range corresponding to the monoisotopic peak of the doubly-charged ion at m/z 819.885 ( m/z window ±5 ppm) for 190 kDa and 170 kDa gel bands acquired in <t>Orbitrap.</t> B : Identification of the ABCA3 tryptic peptide comprising amino acid residues 162–174 of the precursor protein and containing methionine sulfoxide at position 164. MS/MS spectra after CID fragmentation measured in the <t>LTQ</t> ion trap are shown. Major b and y ions are labelled on the spectra and all fragment ions obtained are marked on the identified sequence. The ion at m/z 811 corresponds to the loss of water from the parent ion.
    1100 Ltq Orbitrap Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1100 ltq orbitrap mass spectrometer/product/Thermo Fisher
    Average 85 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    1100 ltq orbitrap mass spectrometer - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

    94
    Thermo Fisher ltq orbitrap elite mass spectrometer
    Rationale and experimental design. A , Confirmation of Δ hik33 growth phenotype under photoautotrophic condition. The deletion of hik33 ), which also contains the photo of the cell culture. B , The WCLs of the WT and Δ hik33 strains of Synechocystis were separated into the membrane (Mem) and the soluble (Sol) fractions. Proteins extracted from either fraction and the WCL were subsequently separated by SDS-PAGE and visualized with Coomassie blue staining. C , Schematic representation of the workflow for the quantitative analysis of the Δ hik33 proteome. Total proteins were extracted from the WCLs of three biological replicates of the WT and Δ hik33 and digested with trypsin. The tryptic peptides were labeled with 6-plex TMT reagents in the order as indicated. The labeled peptides were mixed together with an equal molar ratio, and separated into 15 fractions with RP-HPLC. The peptides in each fraction were quantitatively analyzed by LC-MS/MS using a <t>LTQ-Orbitrap-Elite</t> mass spectrometer.
    Ltq Orbitrap Elite Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1841 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ltq orbitrap elite mass spectrometer/product/Thermo Fisher
    Average 94 stars, based on 1841 article reviews
    Price from $9.99 to $1999.99
    ltq orbitrap elite mass spectrometer - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    85
    Thermo Fisher nanolc ltq orbitrap mass spectrometer
    Rationale and experimental design. A , Confirmation of Δ hik33 growth phenotype under photoautotrophic condition. The deletion of hik33 ), which also contains the photo of the cell culture. B , The WCLs of the WT and Δ hik33 strains of Synechocystis were separated into the membrane (Mem) and the soluble (Sol) fractions. Proteins extracted from either fraction and the WCL were subsequently separated by SDS-PAGE and visualized with Coomassie blue staining. C , Schematic representation of the workflow for the quantitative analysis of the Δ hik33 proteome. Total proteins were extracted from the WCLs of three biological replicates of the WT and Δ hik33 and digested with trypsin. The tryptic peptides were labeled with 6-plex TMT reagents in the order as indicated. The labeled peptides were mixed together with an equal molar ratio, and separated into 15 fractions with RP-HPLC. The peptides in each fraction were quantitatively analyzed by LC-MS/MS using a <t>LTQ-Orbitrap-Elite</t> mass spectrometer.
    Nanolc Ltq Orbitrap Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nanolc ltq orbitrap mass spectrometer/product/Thermo Fisher
    Average 85 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    nanolc ltq orbitrap mass spectrometer - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

    93
    Thermo Fisher ltq orbitrap xltm mass spectrometer
    Rationale and experimental design. A , Confirmation of Δ hik33 growth phenotype under photoautotrophic condition. The deletion of hik33 ), which also contains the photo of the cell culture. B , The WCLs of the WT and Δ hik33 strains of Synechocystis were separated into the membrane (Mem) and the soluble (Sol) fractions. Proteins extracted from either fraction and the WCL were subsequently separated by SDS-PAGE and visualized with Coomassie blue staining. C , Schematic representation of the workflow for the quantitative analysis of the Δ hik33 proteome. Total proteins were extracted from the WCLs of three biological replicates of the WT and Δ hik33 and digested with trypsin. The tryptic peptides were labeled with 6-plex TMT reagents in the order as indicated. The labeled peptides were mixed together with an equal molar ratio, and separated into 15 fractions with RP-HPLC. The peptides in each fraction were quantitatively analyzed by LC-MS/MS using a <t>LTQ-Orbitrap-Elite</t> mass spectrometer.
    Ltq Orbitrap Xltm Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ltq orbitrap xltm mass spectrometer/product/Thermo Fisher
    Average 93 stars, based on 40 article reviews
    Price from $9.99 to $1999.99
    ltq orbitrap xltm mass spectrometer - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    91
    Thermo Fisher ltq velos orbitrap mass spectrometer
    Rationale and experimental design. A , Confirmation of Δ hik33 growth phenotype under photoautotrophic condition. The deletion of hik33 ), which also contains the photo of the cell culture. B , The WCLs of the WT and Δ hik33 strains of Synechocystis were separated into the membrane (Mem) and the soluble (Sol) fractions. Proteins extracted from either fraction and the WCL were subsequently separated by SDS-PAGE and visualized with Coomassie blue staining. C , Schematic representation of the workflow for the quantitative analysis of the Δ hik33 proteome. Total proteins were extracted from the WCLs of three biological replicates of the WT and Δ hik33 and digested with trypsin. The tryptic peptides were labeled with 6-plex TMT reagents in the order as indicated. The labeled peptides were mixed together with an equal molar ratio, and separated into 15 fractions with RP-HPLC. The peptides in each fraction were quantitatively analyzed by LC-MS/MS using a <t>LTQ-Orbitrap-Elite</t> mass spectrometer.
    Ltq Velos Orbitrap Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 524 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ltq velos orbitrap mass spectrometer/product/Thermo Fisher
    Average 91 stars, based on 524 article reviews
    Price from $9.99 to $1999.99
    ltq velos orbitrap mass spectrometer - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    89
    Thermo Fisher ltq orbitrap classic mass spectrometer
    SILAC workflow. A combined strategy involving RNAi against Cdc7, SDS-PAGE protein fractionation, LC–MS/MS data acquisition with a <t>LTQ-Orbitrap</t> (Thermo Fisher Scientific), and downstream bioinformatics for identification, quantification, and functional annotation was applied in this study.
    Ltq Orbitrap Classic Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ltq orbitrap classic mass spectrometer/product/Thermo Fisher
    Average 89 stars, based on 187 article reviews
    Price from $9.99 to $1999.99
    ltq orbitrap classic mass spectrometer - by Bioz Stars, 2020-07
    89/100 stars
      Buy from Supplier

    85
    Thermo Fisher ltq orbitrap etd mass spectrometer
    SILAC workflow. A combined strategy involving RNAi against Cdc7, SDS-PAGE protein fractionation, LC–MS/MS data acquisition with a <t>LTQ-Orbitrap</t> (Thermo Fisher Scientific), and downstream bioinformatics for identification, quantification, and functional annotation was applied in this study.
    Ltq Orbitrap Etd Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ltq orbitrap etd mass spectrometer/product/Thermo Fisher
    Average 85 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    ltq orbitrap etd mass spectrometer - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

    85
    Thermo Fisher ltq orbitrap tandem mass spectrometer
    SILAC workflow. A combined strategy involving RNAi against Cdc7, SDS-PAGE protein fractionation, LC–MS/MS data acquisition with a <t>LTQ-Orbitrap</t> (Thermo Fisher Scientific), and downstream bioinformatics for identification, quantification, and functional annotation was applied in this study.
    Ltq Orbitrap Tandem Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ltq orbitrap tandem mass spectrometer/product/Thermo Fisher
    Average 85 stars, based on 32 article reviews
    Price from $9.99 to $1999.99
    ltq orbitrap tandem mass spectrometer - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

    99
    Thermo Fisher orbitrap ltq orbitrap mass spectrometer
    SILAC workflow. A combined strategy involving RNAi against Cdc7, SDS-PAGE protein fractionation, LC–MS/MS data acquisition with a <t>LTQ-Orbitrap</t> (Thermo Fisher Scientific), and downstream bioinformatics for identification, quantification, and functional annotation was applied in this study.
    Orbitrap Ltq Orbitrap Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/orbitrap ltq orbitrap mass spectrometer/product/Thermo Fisher
    Average 99 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    orbitrap ltq orbitrap mass spectrometer - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    85
    Thermo Fisher ltq orbitrap volos mass spectrometer
    SILAC workflow. A combined strategy involving RNAi against Cdc7, SDS-PAGE protein fractionation, LC–MS/MS data acquisition with a <t>LTQ-Orbitrap</t> (Thermo Fisher Scientific), and downstream bioinformatics for identification, quantification, and functional annotation was applied in this study.
    Ltq Orbitrap Volos Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ltq orbitrap volos mass spectrometer/product/Thermo Fisher
    Average 85 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    ltq orbitrap volos mass spectrometer - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

    93
    Thermo Fisher ltq orbitrap lumos mass spectrometer
    SILAC workflow. A combined strategy involving RNAi against Cdc7, SDS-PAGE protein fractionation, LC–MS/MS data acquisition with a <t>LTQ-Orbitrap</t> (Thermo Fisher Scientific), and downstream bioinformatics for identification, quantification, and functional annotation was applied in this study.
    Ltq Orbitrap Lumos Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ltq orbitrap lumos mass spectrometer/product/Thermo Fisher
    Average 93 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    ltq orbitrap lumos mass spectrometer - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    Image Search Results


    Workflow for MS imaging of Pseudonocardia on the ant propleural plate. a) Photograph and b) cartoon of Pseudonocardia on the ant exoskeleton. c) Grooves were cut into glass slides and double-sided tape was applied to the back of the slide. The ant thoraxes were removed and positioned into the groove of the slide with the propleural plate facing outward and even with the top of the slide. An additional thin strip of double-sided tape was applied below the propleural plate to stabilize and secure the thorax into place. d) Matrix was applied to the slide using an automatic sprayer. e) A laser was fired at the sample to ionize compounds of interest and introduce them into the mass spectrometer. f) An array of mass spectra was acquired using a MALDI-LTQ Orbitrap and g) compiled into MS images. Photo courtesy of Alex Wild.

    Journal: ACS chemical biology

    Article Title: Imaging with Mass Spectrometry of Bacteria on the Exoskeleton of Fungus-Growing Ants

    doi: 10.1021/acschembio.7b00038

    Figure Lengend Snippet: Workflow for MS imaging of Pseudonocardia on the ant propleural plate. a) Photograph and b) cartoon of Pseudonocardia on the ant exoskeleton. c) Grooves were cut into glass slides and double-sided tape was applied to the back of the slide. The ant thoraxes were removed and positioned into the groove of the slide with the propleural plate facing outward and even with the top of the slide. An additional thin strip of double-sided tape was applied below the propleural plate to stabilize and secure the thorax into place. d) Matrix was applied to the slide using an automatic sprayer. e) A laser was fired at the sample to ionize compounds of interest and introduce them into the mass spectrometer. f) An array of mass spectra was acquired using a MALDI-LTQ Orbitrap and g) compiled into MS images. Photo courtesy of Alex Wild.

    Article Snippet: A MALDI- LTQ Orbitrap mass spectrometer (Thermo Scientific) equipped with an N2 laser (spot diameter of 75 μ m) was used in positive ion mode for MSI.

    Techniques: Mass Spectrometry, Imaging, Stripping Membranes, Introduce

    Human Parkin Ser 65 is a substrate of human PINK1 upon CCCP stimulation. ( a ) Confirmation by mass spectrometry that Ser 65 of human Parkin is phosphorylated by CCCP-induced activation of human wild-type PINK1-FLAG. Flp-In T-Rex HEK293 cells expressing FLAG-empty, wild-type PINK1-FLAG, and kinase-inactive PINK1-FLAG (D384A) were co-transfected with HA-Parkin, induced with doxycycline and stimulated with 10 μM of CCCP for 3 h. Whole-cell extracts were obtained following lysis with 1% Triton and approximately 30 mg of whole-cell extract were subjected to immunoprecipitation with anti-HA-agarose and run on 10% SDS-PAGE and stained with colloidal Coomassie blue. Coomassie-stained bands migrating with the expected molecular mass of HA-Parkin were excised from the gel, digested with trypsin, and subjected to high performance liquid chromatography with tandem mass spectrometry (LC-MS-MS) on an LTQ-Orbitrap mass spectrometer. Extracted ion chromatogram analysis of Ser 131 and Ser 65 phosphopeptide (3 + R.NDWTVQNCDLDQQ S IVHIVQRPWR.K+P). The total signal intensity of the phosphopeptide is plotted on the y -axis and retention time is plotted on the x -axis. The m / z value corresponding to the Ser 131 phosphopeptide was detected in all conditions whilst that of the Ser 65 phosphopeptide was only detected in samples from wild-type PINK1-FLAG-expressing cells following CCCP treatment. ( b ) Characterization of Parkin phospho-Ser 65 antibody. Flp-In T-Rex HEK293 cells expressing FLAG-empty, wild-type PINK1-FLAG, and kinase-inactive PINK1-FLAG were co-transfected with untagged wild-type (WT) or Ser 65 Ala (S65A) mutant Parkin, induced with doxycycline and stimulated with 10 μM of CCCP for 3 h. 0.25 mg of 1% Triton whole-cell lysate were subjected to immunoprecipitation with anti-Parkin antibody (S966C) covalently coupled to protein G Sepharose and then immunoblotted with anti-phospho-Ser 65 antibody in the presence of dephosphorylated peptide. Ten per cent of the immunoprecipitate (IP) was immunoblotted with total anti-Parkin antibody. Twenty five micrograms of whole cell lysate was immunoblotted with total PINK1 antibody.

    Journal: Open Biology

    Article Title: PINK1 is activated by mitochondrial membrane potential depolarization and stimulates Parkin E3 ligase activity by phosphorylating Serine 65

    doi: 10.1098/rsob.120080

    Figure Lengend Snippet: Human Parkin Ser 65 is a substrate of human PINK1 upon CCCP stimulation. ( a ) Confirmation by mass spectrometry that Ser 65 of human Parkin is phosphorylated by CCCP-induced activation of human wild-type PINK1-FLAG. Flp-In T-Rex HEK293 cells expressing FLAG-empty, wild-type PINK1-FLAG, and kinase-inactive PINK1-FLAG (D384A) were co-transfected with HA-Parkin, induced with doxycycline and stimulated with 10 μM of CCCP for 3 h. Whole-cell extracts were obtained following lysis with 1% Triton and approximately 30 mg of whole-cell extract were subjected to immunoprecipitation with anti-HA-agarose and run on 10% SDS-PAGE and stained with colloidal Coomassie blue. Coomassie-stained bands migrating with the expected molecular mass of HA-Parkin were excised from the gel, digested with trypsin, and subjected to high performance liquid chromatography with tandem mass spectrometry (LC-MS-MS) on an LTQ-Orbitrap mass spectrometer. Extracted ion chromatogram analysis of Ser 131 and Ser 65 phosphopeptide (3 + R.NDWTVQNCDLDQQ S IVHIVQRPWR.K+P). The total signal intensity of the phosphopeptide is plotted on the y -axis and retention time is plotted on the x -axis. The m / z value corresponding to the Ser 131 phosphopeptide was detected in all conditions whilst that of the Ser 65 phosphopeptide was only detected in samples from wild-type PINK1-FLAG-expressing cells following CCCP treatment. ( b ) Characterization of Parkin phospho-Ser 65 antibody. Flp-In T-Rex HEK293 cells expressing FLAG-empty, wild-type PINK1-FLAG, and kinase-inactive PINK1-FLAG were co-transfected with untagged wild-type (WT) or Ser 65 Ala (S65A) mutant Parkin, induced with doxycycline and stimulated with 10 μM of CCCP for 3 h. 0.25 mg of 1% Triton whole-cell lysate were subjected to immunoprecipitation with anti-Parkin antibody (S966C) covalently coupled to protein G Sepharose and then immunoblotted with anti-phospho-Ser 65 antibody in the presence of dephosphorylated peptide. Ten per cent of the immunoprecipitate (IP) was immunoblotted with total anti-Parkin antibody. Twenty five micrograms of whole cell lysate was immunoblotted with total PINK1 antibody.

    Article Snippet: Isolated phosphopeptides were analysed by LC-MS-MS on a proxeon Easy-nLC nano liquid chromatography system coupled to a Thermo LTQ-orbitrap mass spectrometer.

    Techniques: Mass Spectrometry, Activation Assay, Expressing, Transfection, Lysis, Immunoprecipitation, SDS Page, Staining, High Performance Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Mutagenesis

    (A) Scheme of the methods used to analyze BMSCs Proteins identification was done on an orbitrap LTQ XL with a protein threshold of 99% (FDR 0.1%) and peptide threshold of 97% (FDR 0.1%) and contained at least 2 identified peptides as parameters ( Supplementary data 1 2 ). (B) Label free quantification of immune modulators, neurotrophic and growth factors and apoptotic molecules identified in conditioned media obtained from BMSCs, spinal cord lesion (SCI) or control (non injured) using Scaffold_4.0.6.1. Label-free quantification was done on top3 of the total ion current (TIC). (BMP 1: Bone morphogenic protein 1, Cdh13: T-Cadherin, CLEC11a: C-type lectin domain family 11 member A CTGF: connective tissue growth factor, Epha 3: Ephrin A isoform 3, Epdr1: Ependymin related protein 1, FAM3C: family with sequence similarity 3, member C Gal 1: galectin 1; GAP-43: Growth associated protein 43, Ltbp: Latent TGF-beta binding protein, IgFbp 7: Insulin growth factor binding protein 7, MIF: macrophage inhibiting factor, PDGF: Platelet derived growth factor, PGF: placenta growth factor, spondin 2, SPARC, TIMP-1: Tissue inhibitor metalloproteinase 1 2).

    Journal: Scientific Reports

    Article Title: Modulation properties of factors released by bone marrow stromal cells on activated microglia: an in vitro study

    doi: 10.1038/srep07514

    Figure Lengend Snippet: (A) Scheme of the methods used to analyze BMSCs Proteins identification was done on an orbitrap LTQ XL with a protein threshold of 99% (FDR 0.1%) and peptide threshold of 97% (FDR 0.1%) and contained at least 2 identified peptides as parameters ( Supplementary data 1 2 ). (B) Label free quantification of immune modulators, neurotrophic and growth factors and apoptotic molecules identified in conditioned media obtained from BMSCs, spinal cord lesion (SCI) or control (non injured) using Scaffold_4.0.6.1. Label-free quantification was done on top3 of the total ion current (TIC). (BMP 1: Bone morphogenic protein 1, Cdh13: T-Cadherin, CLEC11a: C-type lectin domain family 11 member A CTGF: connective tissue growth factor, Epha 3: Ephrin A isoform 3, Epdr1: Ependymin related protein 1, FAM3C: family with sequence similarity 3, member C Gal 1: galectin 1; GAP-43: Growth associated protein 43, Ltbp: Latent TGF-beta binding protein, IgFbp 7: Insulin growth factor binding protein 7, MIF: macrophage inhibiting factor, PDGF: Platelet derived growth factor, PGF: placenta growth factor, spondin 2, SPARC, TIMP-1: Tissue inhibitor metalloproteinase 1 2).

    Article Snippet: The chromatography system was coupled to a Thermo Scientific LTQ-Orbitrap XL mass spectrometer programmed to acquire in data-dependent mode.

    Techniques: Sequencing, Binding Assay, Derivative Assay

    Flowchart of the label-free quantification approach. Protein digests of lysates of IECs were prepared from six mice per time point and analyzed in duplicate on the LTQ-Orbitrap, as described in the Experimental Procedures. Raw data sets were processed

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Time-resolved Quantitative Proteome Analysis of In Vivo

    doi: 10.1074/mcp.M110.005231

    Figure Lengend Snippet: Flowchart of the label-free quantification approach. Protein digests of lysates of IECs were prepared from six mice per time point and analyzed in duplicate on the LTQ-Orbitrap, as described in the Experimental Procedures. Raw data sets were processed

    Article Snippet: Protein digests were analyzed in duplicate using a LC-ESI-MS/MS system consisting of a Rheos Allegro pump (Thermo Fisher Scientific, Waltham, MA), a PAL HTC autosampler (CTC Analytics, Zwingen, Switzerland), and an LTQ-XL Orbitrap mass spectrometer (Thermo Fisher Scientific, Waltham, MA) equipped with a nano-ESI source from the same supplier.

    Techniques: Mouse Assay

    Full scan spectra of protein mixtures: MALDI-TOF spectrum of histone mixture, 10 mg/mL (a), HRAM MAI-Orbitrap spectrum of histone mixture, 1 mg/mL (b) and HRAM MAI-Orbitrap spectrum of rat brain protein extract.

    Journal: Analytical and bioanalytical chemistry

    Article Title: Coupling Matrix-Assisted Ionization with High Resolution Mass Spectrometry and Electron Transfer Dissociation to Characterize Intact Proteins and Post-Translational Modifications

    doi: 10.1007/s00216-017-0611-4

    Figure Lengend Snippet: Full scan spectra of protein mixtures: MALDI-TOF spectrum of histone mixture, 10 mg/mL (a), HRAM MAI-Orbitrap spectrum of histone mixture, 1 mg/mL (b) and HRAM MAI-Orbitrap spectrum of rat brain protein extract.

    Article Snippet: In the comparison study, the MALDI and MAI (MALDI) spectra were acquired on MALDI-LTQ-Orbitrap XL mass spectrometer (Thermo Scientific, Bremen, Germany), which is also a hybrid ion-trap-Orbitrap mass spectrometer but with MALDI source.

    Techniques:

    Partial sequence (162–174) is present in unprocessed ABCA3 and missing in processed protein form. A : Intensity profiles of an m/z range corresponding to the monoisotopic peak of the doubly-charged ion at m/z 819.885 ( m/z window ±5 ppm) for 190 kDa and 170 kDa gel bands acquired in Orbitrap. B : Identification of the ABCA3 tryptic peptide comprising amino acid residues 162–174 of the precursor protein and containing methionine sulfoxide at position 164. MS/MS spectra after CID fragmentation measured in the LTQ ion trap are shown. Major b and y ions are labelled on the spectra and all fragment ions obtained are marked on the identified sequence. The ion at m/z 811 corresponds to the loss of water from the parent ion.

    Journal: PLoS ONE

    Article Title: Analysis of the Proteolytic Processing of ABCA3: Identification of Cleavage Site and Involved Proteases

    doi: 10.1371/journal.pone.0152594

    Figure Lengend Snippet: Partial sequence (162–174) is present in unprocessed ABCA3 and missing in processed protein form. A : Intensity profiles of an m/z range corresponding to the monoisotopic peak of the doubly-charged ion at m/z 819.885 ( m/z window ±5 ppm) for 190 kDa and 170 kDa gel bands acquired in Orbitrap. B : Identification of the ABCA3 tryptic peptide comprising amino acid residues 162–174 of the precursor protein and containing methionine sulfoxide at position 164. MS/MS spectra after CID fragmentation measured in the LTQ ion trap are shown. Major b and y ions are labelled on the spectra and all fragment ions obtained are marked on the identified sequence. The ion at m/z 811 corresponds to the loss of water from the parent ion.

    Article Snippet: Mass spectrometric analysis was performed on an LTQ-Orbitrap Discovery mass spectrometer (Thermo Fisher Scientific, Bremen, Germany) operated in a data dependent mode in which each full MS scan (m/z 200–1,450) acquired in Orbitrap with resolution of R = 30,000 (m/z 400) was followed by five MS/MS scans where the five most abundant molecular ions were dynamically selected and fragmented by collision-induced dissociation (CID) using a normalized collision energy of 35% in the LTQ ion trap.

    Techniques: Sequencing, Mass Spectrometry

    Rationale and experimental design. A , Confirmation of Δ hik33 growth phenotype under photoautotrophic condition. The deletion of hik33 ), which also contains the photo of the cell culture. B , The WCLs of the WT and Δ hik33 strains of Synechocystis were separated into the membrane (Mem) and the soluble (Sol) fractions. Proteins extracted from either fraction and the WCL were subsequently separated by SDS-PAGE and visualized with Coomassie blue staining. C , Schematic representation of the workflow for the quantitative analysis of the Δ hik33 proteome. Total proteins were extracted from the WCLs of three biological replicates of the WT and Δ hik33 and digested with trypsin. The tryptic peptides were labeled with 6-plex TMT reagents in the order as indicated. The labeled peptides were mixed together with an equal molar ratio, and separated into 15 fractions with RP-HPLC. The peptides in each fraction were quantitatively analyzed by LC-MS/MS using a LTQ-Orbitrap-Elite mass spectrometer.

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Translating Divergent Environmental Stresses into a Common Proteome Response through the Histidine Kinase 33 (Hik33) in a Model Cyanobacterium *

    doi: 10.1074/mcp.M116.068080

    Figure Lengend Snippet: Rationale and experimental design. A , Confirmation of Δ hik33 growth phenotype under photoautotrophic condition. The deletion of hik33 ), which also contains the photo of the cell culture. B , The WCLs of the WT and Δ hik33 strains of Synechocystis were separated into the membrane (Mem) and the soluble (Sol) fractions. Proteins extracted from either fraction and the WCL were subsequently separated by SDS-PAGE and visualized with Coomassie blue staining. C , Schematic representation of the workflow for the quantitative analysis of the Δ hik33 proteome. Total proteins were extracted from the WCLs of three biological replicates of the WT and Δ hik33 and digested with trypsin. The tryptic peptides were labeled with 6-plex TMT reagents in the order as indicated. The labeled peptides were mixed together with an equal molar ratio, and separated into 15 fractions with RP-HPLC. The peptides in each fraction were quantitatively analyzed by LC-MS/MS using a LTQ-Orbitrap-Elite mass spectrometer.

    Article Snippet: For MS analysis, the peptides were resuspended in 0.1% formic acid (FA) and analyzed by a LTQ Orbitrap Elite mass spectrometer (Thermo Scientific) coupled online to an Easy-nLC 1000 in the data-dependent mode.

    Techniques: Cell Culture, SDS Page, Staining, Labeling, High Performance Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry

    SILAC workflow. A combined strategy involving RNAi against Cdc7, SDS-PAGE protein fractionation, LC–MS/MS data acquisition with a LTQ-Orbitrap (Thermo Fisher Scientific), and downstream bioinformatics for identification, quantification, and functional annotation was applied in this study.

    Journal: Journal of proteome research

    Article Title: Quantitative Proteomics Reveals a “Poised Quiescence” Cellular State after Triggering the DNA Replication Origin Activation Checkpoint

    doi: 10.1021/pr100678k

    Figure Lengend Snippet: SILAC workflow. A combined strategy involving RNAi against Cdc7, SDS-PAGE protein fractionation, LC–MS/MS data acquisition with a LTQ-Orbitrap (Thermo Fisher Scientific), and downstream bioinformatics for identification, quantification, and functional annotation was applied in this study.

    Article Snippet: LC–MS/MS was performed with an LTQ-Orbitrap Classic mass spectrometer (Thermo Fisher Scientific, U.K.).

    Techniques: SDS Page, Fractionation, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Functional Assay