Navigation Links
Complex Peptide Mixture Analysis on the Finnigan LTQ Linear Ion Trap Mass Spectrometer

Chromatography and Mass Spectrometry Application Note

Melissa Chen, Diane Cho, Thermo Electron Corporation, San Jose, CA

Introduction
Rapid identification of low abundance proteins in complex mixtures is one of the key objectives for the use of mass spectrometry in proteomics research. Traditionally, this approach has utilized 3-dimensional quadrupole ion trap systems, such as the Finnigan LCQ Deca XP Plus and the Finnigan ProteomeX. Recent developments in ion trap mass spectrometry, and the introduction of the Finnigan LTQ linear ion trap mass spectrometer provide further tools to boost sample throughput, and produce higher peptide coverage with a higher degree of confidence for protein identification.

Goal
This study demonstrates how the fast cycle time and unparalleled MS/MS sensitivity of the high performance Finnigan LTQ linear ion trap mass spectrometer result in increased coverage, and faster and more confident protein identification.

Experimental
LC/MS/MS analysis of enzymatically modified human plasma samples was performed using NanoSpray ionization. The TurboSEQUEST algorithm was used for data analysis and protein identification.

Sample Preparation
A whole human plasma sample (5 mg, Sigma) was reduced, alkylated, and enzymatically digested. A total of 3 uL of the digested mixture (1 ug/uL) was loaded onto a reversed-phase (RP) C18 column for LC/MS analysis.

LC Separation and MS Analysis
HPLC
HPLC system: Finnigan Surveyor MS pump with a flow splitter.
Column: 0.15100 mm C18 (Thermo Electron)
Flow rate: 600 nL/min
Mobile phase A: Water with 0.1% Formic Acid
B: Acetonitrile with 0.1% Formic Acid

Gradi
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Structural Analysis of Glycosylated Peptides in Complex Mixtures with Ion Trap MSn
2. Purification of the Components of Human Macular Carotenoid-Binding Complex, Rev A
3. Whole Gel Eluter Purification of a Functional Multiprotein DNA Replication Complex, Rev A
4. LC-MALDI and MALDI-MS/MS for the Analysis of Complex Peptide Mixtures
5. Detecting Gastrin-Releasing Peptide Receptor by in situ PCR on Archived Tissue
6. Efficient Primary Structure Elucidation of Disulfide-bridged Peptides
7. Multiplexed Quantitative Peptide Assays for Protein Biomarkers of Cardiovascular Disease in Human Plasma
8. Direct Monitoring of Solid-Phase Peptide Synthesis by MALDI-TOF MS
9. Separation of Peptides by Hydrophilic Interaction Chromatography Using TSKgel Amide-80
10. High-Fidelity PCR with a Novel Polymerase Mixture
11. High Electrotransformation Efficiencies Obtained With DNA From Ligation Mixtures
Post Your Comments:
(Date:9/8/2008)... Funds to Support Clinical Development of Carfi...d of Directors, SOUTH SAN FRANCISCO, Calif., Sept...he discovery and development of novel therapeutics...r and autoimmune diseases, today,announced the suc...,million. The Series C round was led by new invest...
(Date:9/8/2008)... Symphony Management Consulting a...tions Director. , Charlot...consulting.com [Symphony Management Consulting] a...tions Director., , , , ,Mr. Whalen joins Symphony...e was responsible for designing, developing, and p...
(Date:9/5/2008)...ASSAS, Va. -- With the publication of a study led ...University,s Department of Molecular and Microbiol...nderstanding how the human immunodeficiency virus ...ich is caused by HIV, affected more than 33 millio...th Organization statistics. , In the Sept. 5 iss...
(Date:9/5/2008)...terials have exercised as much of a hold on the hu... , But for all of its popular uses money, medal...t lay hidden until the 1980s, when Masatake Haruta...t gold, which had long been considered inactive, c...emonstrated the low-temperature oxidation of CO an...
Breaking Biology Technology:Proteolix Raises $79 Million in Series C Financing 2Proteolix Raises $79 Million in Series C Financing 3Symphony Managemnt Consulting Hires New Solutions Director 2Researchers decode viral process that prepares cells for HIV infection 2Scientists peel away the mystery behind gold's catalytic prowess 2Scientists peel away the mystery behind gold's catalytic prowess 3Scientists peel away the mystery behind gold's catalytic prowess 4
...ssue storage , solution that st... tissues. (Ambion,s new RNA later -I...or use with frozen tissues - see "<A , .../1.html">Stop , Grinding Frozen... harvested and immediately submerged in ...
...GATE THIS ARTICLE: <A , hre... href="#3">RNA , Isolation, Probe...lectrophoresis, , Transfer > <A , ... Detection > <A , href="... , , , Northe...
...ed with fluorescent , dyes (i.e. Cy3 ... directly incorporated during cDNA synth...aRNA reverse transcription (RT) reaction, or attac...n performed after any enzymatic reactions , ...the enzymatic , incorporation of a Cy...
...R on Sympathetic Neuron Lysates , , Silenci...ated siRNAs , , D...ron Lysates , Quantitative RT-PCR is t...r measuring mRNA levels from small numbers of cell...be difficult when cell samples are , ...
Other Biology Technology:RNArticles: Keeping RNA Intact Prior to RNA Isolation - RNAlater 2RNArticles: Keeping RNA Intact Prior to RNA Isolation - RNAlater 3RNArticles: Keeping RNA Intact Prior to RNA Isolation - RNAlater 4RNArticles: Keeping RNA Intact Prior to RNA Isolation - RNAlater 5RNArticles: Keeping RNA Intact Prior to RNA Isolation - RNAlater 6RNArticles: Keeping RNA Intact Prior to RNA Isolation - RNAlater 7Northern Analysis: The Basics 2Northern Analysis: The Basics 3Northern Analysis: The Basics 4Northern Analysis: The Basics 5Northern Analysis: The Basics 6Northern Analysis: The Basics 7Northern Analysis: The Basics 8Northern Analysis: The Basics 9Northern Analysis: The Basics 10Northern Analysis: The Basics 11Northern Analysis: The Basics 12Northern Analysis: The Basics 13Northern Analysis: The Basics 14Northern Analysis: The Basics 15Northern Analysis: The Basics 16Northern Analysis: The Basics 17Northern Analysis: The Basics 18Northern Analysis: The Basics 19Amino Allyl Labeling for Array Analysis 2YOUR DATA: , Direct Real-time RT-PCR on Sympathetic Neuron Lysates, , Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs 2YOUR DATA: , Direct Real-time RT-PCR on Sympathetic Neuron Lysates, , Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs 3YOUR DATA: , Direct Real-time RT-PCR on Sympathetic Neuron Lysates, , Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs 4YOUR DATA: , Direct Real-time RT-PCR on Sympathetic Neuron Lysates, , Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs 5
(Date:9/7/2008)... In a study published by the Lancet journal tod... that a new once-weekly treatment for type 2 diabe...ction. , "Over two million Canadians have diabet...and Senior Investigator at the Samuel Lunenfeld Re...s currently no available therapy for type 2 diabet...
(Date:9/5/2008)...iety is pleased to announce the recipients of its ...eive their awards at the Society,s 53rd Annual Mee...tion and Exhibitions Center in Massachusetts. The ... of California, San Francisco, will receive the An...contributions to transmembrane biology, and pionee...
(Date:9/5/2008)... face the challenge of surviving in a changing cli...niversit Claude Bernard Lyon have shown how a shee... of Scotland responds to two consequences of clima...dictability of winter storms. Dr. Thomas Ezard, le...good and food is plentiful, lambs contribute almos...
(Date:9/5/2008)...ave evolved more than once, according to research ...ondon scientists. , Species of bat with the abil... evolutionary tree of life - some are more related.... This has raised the question of whether echoloca...r the fruit bats somehow lost their ability to ech...
Breaking Biology News(10 mins):New once-a-week treatment for type 2 diabetes developed by Mount Sinai researcher 2Biophysical Society names 2009 award recipients 2Molecular evolution is echoed in bat ears 2Turned off cannabinoid receptor turns on colorectal tumor growth 24841 1Turned off cannabinoid receptor turns on colorectal tumor growth 24841 2Turned off cannabinoid receptor turns on colorectal tumor growth 24841 3Lab study shows methadone breaks resistance in untreatable forms of leukemia 4303 1Lab study shows methadone breaks resistance in untreatable forms of leukemia 4303 2Mayo Clinic on the Summer Olympic Games 3A Better Athletes the Physiology of Performance the Lactate Threshold the Aging Athlete and Much More 24837 1Mayo Clinic on the Summer Olympic Games 3A Better Athletes the Physiology of Performance the Lactate Threshold the Aging Athlete and Much More 24837 2Mayo Clinic on the Summer Olympic Games 3A Better Athletes the Physiology of Performance the Lactate Threshold the Aging Athlete and Much More 24837 3Mayo Clinic on the Summer Olympic Games 3A Better Athletes the Physiology of Performance the Lactate Threshold the Aging Athlete and Much More 24837 4Mayo Clinic on the Summer Olympic Games 3A Better Athletes the Physiology of Performance the Lactate Threshold the Aging Athlete and Much More 24837 5Mayo Clinic on the Summer Olympic Games 3A Better Athletes the Physiology of Performance the Lactate Threshold the Aging Athlete and Much More 24837 6Mayo Clinic on the Summer Olympic Games 3A Better Athletes the Physiology of Performance the Lactate Threshold the Aging Athlete and Much More 24837 7Free articles get read but dont generate more citations 24832 1Free articles get read but dont generate more citations 24832 2
...ervices University of the Health Sciences (USU) su...e that protects against the myriad strains of the ...ive vaccine may need to stimulate the body to prod... multiple viral strains. These results demonstrate...ng antibodies may be an achievable goal. This grou...
...chronic lung disease typically characterized by th...h or cough. Most patients live about five years af...eing published today in the online journal PLoS ON...ofile has a much more rapid progression to complet...plant. , Based on observations in the clinic that ...
...ing out for other mothers, kids. , An intriguing...y the University of Alberta in Edmonton, Canada an...owed that both species responded to the recorded d...licited when coyotes attack fawns, with mule deer ... calls, even when their own fawn stood next to the...
...ctive method for revealing anatomical details of s...e images even clearer and allow physiological proc...dolinium complexes currently used as MRI contrast ...ed in the journal Angewandte Chemie, Korean resear...r and Taeghwan Hyeon at Seoul National University ...
Other Biology News:Gene expression patterns predict rapid decline in idiopathic pulmonary fibrosis patients 2Mule deer moms rescue other fawns 2
The SVT3-2 diverter valve is a 3-port, 2-position valve for use with the BioLogic DuoFlow system.
4',6-Diamidino-2-phenylindole, dihydrochloride (DAPI)
...ations. The cell has a six-gel capacity with effic...-cell can be customized for several parameters: ge...system. The PROTEAN II xi multi-cell can also be c...er format that accommodates 17 and 18 cm IPG strip...
...very process in mind. Due to its in-built flexibil...eal instrument to bridge the gap between assay dev...a fully modular monochromator-based microplate det...tection techniques. In assay development, which re...
Biology Products: