Multiwell in-gel protein digestion and microscale sample preparation for protein identification by mass spectrometry.

Abstract:

:In-gel peptide digestion has become a widely used technique for characterizing proteins resolved by two-dimensional gel electrophoresis. Peptides generated from gel pieces are frequently contaminated with detergent and salts. Prior to matrix-assisted laser desorption/ionization-time of flight mass spectrometry analysis, these contaminants are removed using micro scale C18 sample preparation columns. In this paper, data are presented to demonstrate the application of a solvent resistant MultiScreen 96-well plate with a low peptide binding membrane and ZipTip micropipette based sample preparation. Recoveries of peptides (m/z of 1000 to 5000 Da) derived from standard protein protease digests, were estimated at various stages of the analytical process. An optimized protocol has been established and all the reagents and consumables have been packaged in a ready to use commercial kit. Data will be presented to show the application of this technology package to accelerate the throughput of protein characterization by protease fragmentation.

journal_name

Proteomics

journal_title

Proteomics

authors

Pluskal MG,Bogdanova A,Lopez M,Gutierrez S,Pitt AM

doi

10.1002/1615-9861(200202)2:2<145::aid-prot145>3.0.

subject

Has Abstract

pub_date

2002-02-01 00:00:00

pages

145-50

issue

2

eissn

1615-9853

issn

1615-9861

pii

10.1002/1615-9861(200202)2:2<145::AID-PROT145>3.0.

journal_volume

2

pub_type

杂志文章
  • Apoptosis induced by staurosporine alters chaperone and endoplasmic reticulum proteins: Identification by quantitative proteomics.

    abstract::Apoptosis contributes to cell death after cerebral ischaemia. A quantitative proteomics approach has been employed to define alterations in protein levels in apoptosis induced with staurosporine (STS). Human neuroblastoma derived SH-SY5Y cells were treated with STS (500 nM for 6 h) to induce apoptosis. Quantitative 2-...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200600964

    authors: Short DM,Heron ID,Birse-Archbold JL,Kerr LE,Sharkey J,McCulloch J

    更新日期:2007-09-01 00:00:00

  • Integration of mass spectrometry and RNA-Seq data to confirm human ab initio predicted genes and lncRNAs.

    abstract::MS/MS has been used to improve genome annotation in various organisms. The classical approach is to construct comprehensive theoretical peptide database with six frame translation model from the whole ORF of a genome and search against this database with real MS/MS spectra. In this work we took a more focused approach...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201400174

    authors: Sun H,Chen C,Shi M,Wang D,Liu M,Li D,Yang P,Li Y,Xie L

    更新日期:2014-12-01 00:00:00

  • A simple and inexpensive approach to interfacing high-performance liquid chromatography and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry.

    abstract::The ability to obtain the accurate mass of a protein in a complex sample mixture aids in determining its correct in vivo form. This is important when identifying post-translationally modified proteins, protein variants or isoforms. The central technique used to separate proteins, 2-dimensional gel electrophoresis offe...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200300843

    authors: Fung KY,Askovic S,Basile F,Duncan MW

    更新日期:2004-10-01 00:00:00

  • Cleavage and functional loss of human apolipoprotein E by digestion of matrix metalloproteinase-14.

    abstract::By means of a degradomic approach applying proteomic techniques, we previously suggested that apolipoprotein E (apoE) is a substrate of matrix metalloproteinase-14 (MMP-14). Here we confirm that apoE is, in fact, a substrate of MMP-14 and also of MMP-7 and MMP-2 to a lesser extent. The 34 kDa apoE protein was initiall...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200700487

    authors: Park JH,Park SM,Park SH,Cho KH,Lee ST

    更新日期:2008-07-01 00:00:00

  • Identification of caspase-3 degradome by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight analysis.

    abstract::The activation of caspases is a critical event for the execution phase of programmed cell death. Caspases are highly specific in their ability to activate or inhibit many crucial proteins in the cell via site-specific cleavage. To date, more than 60 proteins have been shown to be substrates of one or more caspases in ...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200400979

    authors: Lee AY,Park BC,Jang M,Cho S,Lee DH,Lee SC,Myung PK,Park SG

    更新日期:2004-11-01 00:00:00

  • Quantitative proteome analysis of age-related changes in mouse gastrocnemius muscle using mTRAQ.

    abstract::Aging is associated with a progressive loss of skeletal muscular function that often leads to progressive disability and loss of independence. Although muscle aging is well documented, the molecular mechanisms of this condition still remain unclear. To gain greater insight into the changes associated with aging of ske...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201200497

    authors: Hwang CY,Kim K,Choi JY,Bahn YJ,Lee SM,Kim YK,Lee C,Kwon KS

    更新日期:2014-01-01 00:00:00

  • Modification-specific proteomics: strategies for characterization of post-translational modifications using enrichment techniques.

    abstract::More than 300 different types of protein post-translational modifications (PTMs) have been described, many of which are known to have pivotal roles in cellular physiology and disease. Nevertheless, only a handful of PTMs have been extensively investigated at the proteome level. Knowledge of protein substrates and thei...

    journal_title:Proteomics

    pub_type: 杂志文章,评审

    doi:10.1002/pmic.200900398

    authors: Zhao Y,Jensen ON

    更新日期:2009-10-01 00:00:00

  • Improving gel-based proteome analysis of soluble protein extracts by heat prefractionation.

    abstract::The presence of high-abundance proteins in complex protein mixtures often masks low-abundance proteins and causes loss of resolution of 2DE. Protein fractionation steps conducted prior to 2DE can enhance the detection of low-abundance proteins and improve the resolution of 2DE. Here, we report a method to prefractiona...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201100475

    authors: Wang W,Wu X,Xiong E,Tai F

    更新日期:2012-04-01 00:00:00

  • In vivo proteome dynamics during early bovine myogenesis.

    abstract::Myogenesis is a complex process of which the underlying mechanisms are conserved between species, including birds and mammals. Despite a good understanding of the stages of myogenesis, many of the mechanisms involved in the regulation of proliferation of the successive myoblast generations, the cellular transitions ce...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200701101

    authors: Chaze T,Meunier B,Chambon C,Jurie C,Picard B

    更新日期:2008-10-01 00:00:00

  • Proteomic analysis of the anterior cingulate cortex in the major psychiatric disorders: Evidence for disease-associated changes.

    abstract::Abnormalities of the anterior cingulate cortex have previously been described in schizophrenia, major depressive disorder and bipolar disorder. In this study 2-DE was performed followed by mass spectrometric sequencing to identify disease-specific protein changes within the anterior cingulate cortex in these psychiatr...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200500069

    authors: Beasley CL,Pennington K,Behan A,Wait R,Dunn MJ,Cotter D

    更新日期:2006-06-01 00:00:00

  • An efficient solubilization buffer for plant proteins focused in immobilized pH gradients.

    abstract::The solubilization of a large array of proteins before electrophoresis itself is a very critical point for proteomic analyses. We compared the efficiency of several different solubilization buffers. From this work, we defined a very efficient solubilization buffer, including two chaotropes, two reducing agents (R2), t...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200300450

    authors: Méchin V,Consoli L,Le Guilloux M,Damerval C

    更新日期:2003-07-01 00:00:00

  • Gel-free analysis of the human brain proteome: application of liquid chromatography and mass spectrometry on biopsy and autopsy samples.

    abstract::This paper reports on the findings of the Biomedical Research Institute, as one of the participants in the pilot study of the HUPO Brain Proteome Project. A biopsy and autopsy study sample derived from human brain was distributed among the participants for proteomic analysis. In our laboratory, attention was focused o...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200600080

    authors: Dumont D,Noben JP,Verhaert P,Stinissen P,Robben J

    更新日期:2006-09-01 00:00:00

  • Differential expression of the liver proteome in senescence accelerated mice.

    abstract::The senescence-accelerated mouse (SAM) is a useful animal model to study aging or age-associated disorders due to its inherited aging phenotype. To investigate proteins involved in the aging process in liver, we compared the young (4- or 20-week old) and the aged group (50-week-old) of SAMP8 (short life span) and SAMR...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200300562

    authors: Cho YM,Bae SH,Choi BK,Cho SY,Song CW,Yoo JK,Paik YK

    更新日期:2003-10-01 00:00:00

  • Placenta Stem/Stromal Cell-Derived Extracellular Vesicles for Potential Use in Lung Repair.

    abstract::Many acute and chronic lung injuries are incurable and rank as the fourth leading cause of death globally. While stem cell treatment for lung injuries is a promising approach, there is growing evidence that the therapeutic efficacy of stem cells originates from secreted extracellular vesicles (EVs). Consequently, EVs ...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201800166

    authors: Kim SY,Joglekar MV,Hardikar AA,Phan TH,Khanal D,Tharkar P,Limantoro C,Johnson J,Kalionis B,Chrzanowski W

    更新日期:2019-09-01 00:00:00

  • The methylproteome and the intracellular methylation network.

    abstract::Since its discovery more than 50 years ago, post-translational modification (PTM) of proteins via methylation has grown in prominence, its involvement having been recognised in a number of central processes in the cell. Of these, the best characterised is its role in the epigenetic code. However, there is increasing e...

    journal_title:Proteomics

    pub_type: 杂志文章,评审

    doi:10.1002/pmic.201100397

    authors: Erce MA,Pang CN,Hart-Smith G,Wilkins MR

    更新日期:2012-02-01 00:00:00

  • A detergent- and cyanogen bromide-free method for integral membrane proteomics: application to Halobacterium purple membranes and the human epidermal membrane proteome.

    abstract::A simple and rapid method for characterizing hydrophobic integral membrane proteins and its utility for membrane proteomics using microcapillary liquid chromatography coupled on-line with tandem mass spectrometry (microLC-MS/MS) is described. The present technique does not rely on the use of detergents, strong organic...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200300543

    authors: Blonder J,Conrads TP,Yu LR,Terunuma A,Janini GM,Issaq HJ,Vogel JC,Veenstra TD

    更新日期:2004-01-01 00:00:00

  • A proteomic approach to cisplatin resistance in the cervix squamous cell carcinoma cell line A431.

    abstract::Since drug resistance is a complex and multifactorial event involving activation/repression of multiple biochemical pathways, we used a proteomic approach to study cisplatin resistance and drug response in human tumor cell lines. The cervix squamous cell carcinoma cell line A431 and its cisplatin-resistant subline, A4...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200400835

    authors: Castagna A,Antonioli P,Astner H,Hamdan M,Righetti SC,Perego P,Zunino F,Righetti PG

    更新日期:2004-10-01 00:00:00

  • Radioresistance-related proteins in rectal cancer.

    abstract::Radiation therapy (RT), combined with chemotherapy, is currently the standard adjuvant approach for UICC-stage II and III rectal cancer patients. Individual rectal tumors display wide ranges of radiosensitivity (RS). The aim of the present study was to identify proteins associated with radioresistance (RR), with the f...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200300854

    authors: Allal AS,Kähne T,Reverdin AK,Lippert H,Schlegel W,Reymond MA

    更新日期:2004-08-01 00:00:00

  • GlycoMod--a software tool for determining glycosylation compositions from mass spectrometric data.

    abstract::GlycoMod (http://www.expasy.ch/tools/glycomod/) is a software tool designed to find all possible compositions of a glycan structure from its experimentally determined mass. The program can be used to predict the composition of any glycoprotein-derived oligosaccharide comprised of either underivatised, methylated or ac...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/1615-9861(200102)1:2<340::AID-PROT340>3.0.

    authors: Cooper CA,Gasteiger E,Packer NH

    更新日期:2001-02-01 00:00:00

  • An improved method of sample preparation on AnchorChip targets for MALDI-MS and MS/MS and its application in the liver proteome project.

    abstract::An improved method for sample preparation for MALDI-MS and MS/MS using AnchorChip targets is presented. The method, termed the SMW method (sample, matrix wash), results in better sensitivity for peptide mass fingerprinting as well as for sequencing by MS/MS than previously published methods. The method allows up-conce...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200600184

    authors: Zhang X,Shi L,Shu S,Wang Y,Zhao K,Xu N,Liu S,Roepstorff P

    更新日期:2007-07-01 00:00:00

  • 2-D DIGE analysis of the proteome of extracts from peanut variants reveals striking differences in major allergen contents.

    abstract::Over the last decade, an increasing prevalence of peanut allergies was observed worldwide. Peanuts are meanwhile categorized among the most dangerous food allergens. This is particularly relevant since peanut-derived ingredients are widely used in industrial food production. To minimize the problem of hidden food alle...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200800938

    authors: Schmidt H,Gelhaus C,Latendorf T,Nebendahl M,Petersen A,Krause S,Leippe M,Becker WM,Janssen O

    更新日期:2009-07-01 00:00:00

  • Mass spectrometry-based immuno-precipitation proteomics - the user's guide.

    abstract::Immuno-precipitation (IP) experiments using MS provide a sensitive and accurate way of characterising protein complexes and their response to regulatory mechanisms. Differences in stoichiometry can be determined as well as the reliable identification of specific binding partners. The quality control of IP and protein ...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201000548

    authors: ten Have S,Boulon S,Ahmad Y,Lamond AI

    更新日期:2011-03-01 00:00:00

  • Mapping protein cysteine sulfonic acid modifications with specific enrichment and mass spectrometry: an integrated approach to explore the cysteine oxidation.

    abstract::Oxidation of thiol proteins, which results in conversion of cysteine residues to cysteine sulfenic, sulfinic or sulfonic acids, is an important posttranslational control of protein function in cells. To facilitate the analysis of this process with MALDI-MS, we have developed a method for selective enrichment and ident...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200900850

    authors: Chang YC,Huang CN,Lin CH,Chang HC,Wu CC

    更新日期:2010-08-01 00:00:00

  • Surface plasmon resonance imaging protein arrays for analysis of triple protein interactions of HPV, E6, E6AP, and p53.

    abstract::We have developed a surface plasmon resonance (SPR)-based protein microarray to study protein-protein interactions in a high-throughput mode. As a model system, triple protein interactions have been explored with human papillomaviral E6 protein, tumor suppressor p53, and ubiquitin ligase E6AP. Human papillomavirus (HP...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200500635

    authors: Ro HS,Koh BH,Jung SO,Park HK,Shin YB,Kim MG,Chung BH

    更新日期:2006-04-01 00:00:00

  • From spots to beads-PTM-peptide bead arrays for the characterization of anti-histone antibodies.

    abstract::Antibodies that recognize PTMs of histones play a central role in epigenetic proteomic research. Modification-specific antibodies are employed in chromatin immunoprecipitation, for Western blotting and during the immunoprecipitation steps for MS-based global proteomic analyses. Knowledge about the antibodies' off-targ...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201200383

    authors: Heubach Y,Planatscher H,Sommersdorf C,Maisch D,Maier J,Joos TO,Templin MF,Poetz O

    更新日期:2013-03-01 00:00:00

  • Human plasma proteome analysis by reversed sequence database search and molecular weight correlation based on a bacterial proteome analysis.

    abstract::In shotgun proteomics, proteins can be fractionated by 1-D gel electrophoresis and digested into peptides, followed by liquid chromatography to separate the peptide mixture. Mass spectrometry generates hundreds of thousands of tandem mass spectra from these fractions, and proteins are identified by database searching....

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200500318

    authors: Park GW,Kwon KH,Kim JY,Lee JH,Yun SH,Kim SI,Park YM,Cho SY,Paik YK,Yoo JS

    更新日期:2006-02-01 00:00:00

  • Chemoenzymatic method for glycomics: Isolation, identification, and quantitation.

    abstract::Over the past decade, considerable progress has been made with respect to the analytical methods for analysis of glycans from biological sources. Regardless of the specific methods that are used, glycan analysis includes isolation, identification, and quantitation. Derivatization is indispensable to increase their ide...

    journal_title:Proteomics

    pub_type: 杂志文章,评审

    doi:10.1002/pmic.201500266

    authors: Yang S,Rubin A,Eshghi ST,Zhang H

    更新日期:2016-01-01 00:00:00

  • Proteomic analysis of the phytopathogenic fungus Botrytis cinerea during cellulose degradation.

    abstract::The ascomycete Botrytis cinerea is a phytopathogenic fungus infecting and causing significant yield losses in a number of crops. Moreover, in the last few years, B. cinerea has been adopted as an important model system in molecular phytopathology. In spite of these contributions, the molecular basis of the infection c...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200800540

    authors: Fernández-Acero FJ,Colby T,Harzen A,Cantoral JM,Schmidt J

    更新日期:2009-05-01 00:00:00

  • Lipid particle composition of the yeast Yarrowia lipolytica depends on the carbon source.

    abstract::Lipid particles (LP) of all types of cells are a depot of neutral lipids. The present investigation deals with the isolation of LP from the yeast Yarrowia lipolytica and the characterization of their lipid and protein composition. Properties of LP varied depending on the carbon source. LP from glucose-grown cells reve...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200500339

    authors: Athenstaedt K,Jolivet P,Boulard C,Zivy M,Negroni L,Nicaud JM,Chardot T

    更新日期:2006-03-01 00:00:00

  • Proteomic expression analysis of colorectal cancer by two-dimensional differential gel electrophoresis.

    abstract::The identification of specific protein markers for colorectal cancer would provide the basis for early diagnosis and detection, as well as clues for understanding the molecular mechanisms governing cancer progression. In this report, we describe the proteomic analysis of the samples of colorectal cancer corresponding ...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200401196

    authors: Alfonso P,Núñez A,Madoz-Gurpide J,Lombardia L,Sánchez L,Casal JI

    更新日期:2005-07-01 00:00:00