Navigation Links
2D-LC analysis of phosphopeptides in brain tissue using Ettan MDLC and Finnigan LTQ

Key words: phosphopeptide • two-dimensional liquid chromatography (2D-LC) • Ettan MDLC • LC-MS/MS • reversed-phase chromatography (RPC) • strong cation exchange (SCX) chromatography • linear ion trap mass spectrometer


A 2D–LC-MS method was developed to analyze phospho-peptides in mouse brain tissue. The trypsin-digested tissue was separated by strong cation exchange chromatography (SCX), followed by reversed-phase chromatography (RPC) using Ettan™ MDLC. The detection was performed by mass spectrometry using neutral loss of phosphoric acid to selectively detect the phosphorylated peptides. Several phosphorylation sites were found, and a strategy for confident assignment of these was developed.


Introduction
One of the most important post-translational modifications is phosphorylation of serine, threonine, or tyrosine residues. Phosphorylated proteins play important roles in a wide range of biological processes, such as signal transduction, apoptosis, and cell cycle control. Detection of phosphorylation sites by mass spectrometry in proteins extracted from biological material is hampered by the low abundance, low stoichiometry, and poor ionization of phosphopeptides (1).

In this work, a biocompatible nanoscale liquid chromatography (LC) system, Ettan MDLC, was used for separating phosphopeptides. No metal ions that can chelate phosphate groups are present in the fluid pathway of the LC system, resulting in highly sensitive analyses (2).

Separation of the tryptic peptides was performed in two dimensions, SCX followed by RPC. A Finnigan™ LTQ™ linear ion trap mass spectrometer was used for detecting phosphopeptides by fragmenting all peptides that exhibited a neutral loss of phosphoric acid.


Materials
Products used
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. High-throughput gene expression analysis to screen for anti-asthma drugs
2. Fully automated DNA purification and efficient multiplex PCR for analysis of microsatellite loci
3. Flow cytometric analysis of human primary cells using the Agilent 2100 bioanalyzer and on-chip staining
4. 2100 expert software Powerful software for the analysis of RNA, DNA, proteins, and cells with the Agilent 2100 bioanalyzer
5. Using the Agilent 2100 bioanalyzer for quality control of protein samples prior to MS-analysis
6. Microfluidic analysis of multiplex PCR products for the genotyping of Helicobacter pylori
7. High resolution DNA analysis with the DNA 500 and DNA 1000 LabChip kits
8. Extended data analysis using DeCyder EDA
9. Neurite outgrowth cell-by-cell analysis using the IN Cell Developer Toolbox
10. Laser Capture Microdissection of muscle fiber populations and expression analysis by RT-PCR
11. TSKgel ODS-100Z and TSKgel ODS-100V column series provides a universal solution for the analysis of samples containing acidic, basic, polar & non-polar compounds
Post Your Comments:
(Date:8/19/2014)... Aug. 19, 2014 A Geisinger researcher on ... for a Centers for Disease Control and Prevention (CDC) ... public health concern. Joseph Boscarino , ... of a team of researchers nominated for the Centers ... Science Award for Data Methods and Study Design. ...
(Date:8/19/2014)... 19, 2014 Research and Markets  has announced ... - Global Strategic Business Report" report to their offering. ... Solid Oxide Fuel Cells (SOFCs) in US$ Thousands. The report ... , Europe , Asia-Pacific ... are provided for the period 2012 through 2020. Market data ...
(Date:8/19/2014)... Lenexa, KS (PRWEB) August 19, 2014 ... to offer fast and cost-effective metabolite production from ... and proprietary chemistry platform, the technology enables convenient ... issues. According to XenoTech’s Vice President of ... metabolite synthesis for assessing clinical risk can be ...
(Date:8/19/2014)... Aug. 19, 2014 CSSi, the leader in ... announced the formation of the company,s Medical and Clinical ... addition of Dr. William E. Gannon, Jr. ... The MCAB, with Dr. Gannon,s ... therapeutic areas and set strategic goals for the advancement ...
Breaking Biology Technology:Geisinger investigator nominated for federal award 2Geisinger investigator nominated for federal award 3Global Solid Oxide Fuel Cells (SOFCs) - Strategic Business Report 2014 2Beyond Metabolite Identification, Testable Quantities Now Offered by XenoTech 2Beyond Metabolite Identification, Testable Quantities Now Offered by XenoTech 3CSSi Appoints Dr. William E. Gannon, Jr. to Newly Formed Medical and Clinical Advisory Board 2CSSi Appoints Dr. William E. Gannon, Jr. to Newly Formed Medical and Clinical Advisory Board 3
... Sept. 1 Express Scripts, Inc. (Nasdaq: ESRX ) announced that ... Thomas Weisel Partners Healthcare Conference 2009 on September 9, 2009 at ... , Morgan Stanley Global Healthcare Conference on September 15, 2009 at ... , These presentations will also be broadcast via the Internet ...
... PHILADELPHIA, Sept. 1 ERT (Nasdaq: ERES ), announced ... Keith Schneck, the Company,s Executive Vice President and CFO, are scheduled to ... on September 9, 2009 in New York City, New York. , ... Company,s corporate website at http://www.ert.com . The webcast may ...
... Sept. 1 Environmental Tectonics Corporation,s (OTC Bulletin Board: ETCC) ... sale of a BARA-MED((R)) XD Monoplace Hyperbaric Chamber to ... Georgia area. , , Hyperbaric Oxygen ... the patient breathes 100% oxygen while fully enclosed in a ...
Cached Biology Technology:Express Scripts to Present at Investor Conferences 2ERT to Present at Baird's 2009 Health Care Conference on September 9, 2009 2Environmental Tectonics Corporation's BioMedical Systems Division Announces the Sale of a Monoplace Hyperbaric Chamber 2Environmental Tectonics Corporation's BioMedical Systems Division Announces the Sale of a Monoplace Hyperbaric Chamber 3Environmental Tectonics Corporation's BioMedical Systems Division Announces the Sale of a Monoplace Hyperbaric Chamber 4
(Date:8/20/2014)... 20, 2014 /CNW/ -  Issue Serious ... quickly become life threatening for some children.    ... million Canadians have food allergies and this number may be ... children are estimated to have food allergies.  ... irritations and hives to breathing difficulties and loss of consciousness. ...
(Date:8/20/2014)... State University, the Wellcome Trust Sanger Institute and the ... Mycobacterium pinnipedii from skeletons found in Peru ... is a relative of the TB bacterium that affects ... These researchers assume that seals carried the pathogens from ... lions was unexpected" comments Sebastien Gagneux, from the Swiss ...
(Date:8/20/2014)... image of the Para and Mato Grosso states of ... set in order to deforest the land. Deforestation is ... where the land is thereafter converted to a nonforest ... farms, ranches, or urban use. The herringbone-patterned tan lines ... in the middle of the image are evidence of ...
Breaking Biology News(10 mins):Information Update - Allergens and food safety at school - What's in your child's lunch? 2Information Update - Allergens and food safety at school - What's in your child's lunch? 3
... MARC (Maximizing Access to Research Careers) Program has announced ... Summer Research Opportunity Program (SROP). The FASEB MARC Program ... underrepresented groups for summer research experiences in the laboratories ... mentors for the students. The FASEB MARC SROP provides ...
... researchers should welcome the news from the Lawrence Berkeley ... scientists has discovered a new and possibly more effective ... implications for advanced biofuels and therapeutic drugs, as genetically ... to play a key role in the green chemistry ...
... could make more tomatoes taste like heirlooms, reports an international ... scientist. The finding, which will be reported in the ... significant implications for the U.S. tomato industry, which annually harvests ... and fresh-market sales. "This information about the gene responsible ...
Cached Biology News:Undergraduate Student Summer Research Award recipients announced for the 2012 FASEB MARC summer research opportunity program 2Undergraduate Student Summer Research Award recipients announced for the 2012 FASEB MARC summer research opportunity program 3Undergraduate Student Summer Research Award recipients announced for the 2012 FASEB MARC summer research opportunity program 4Programmable DNA scissors found for bacterial immune system 2Programmable DNA scissors found for bacterial immune system 3Discovery may lead to new tomato varieties with vintage flavor and quality 2
Mouse monoclonal antibody raised against a partial recombinant L3MBTL2. NCBI Entrez Gene ID = L3MBTL2...
Mouse monoclonal antibody raised against a partial recombinant QARS. NCBI Entrez Gene ID = QARS...
96-well format (For processing samples on microplate-size MALDI target plates such as those from Bruker, Shimadzu, Thermo, and PerkinElmer) Packaging: 2 / pack...
... The FUNNY SHAKERS are multi-functional mixing ... This device is a solid, heavy-duty durable ... is both mixing and vortexing in nature. ... based keypad with digital display to control ...
Biology Products: