Navigation Links
LC-MALDI and MALDI-MS/MS for the Analysis of Complex Peptide Mixtures

).



Methods
Protein Mixture: A 50 protein mixture (5 pmol of each protein) was digested using trypsin. 5 uL of the digest supernatent (500 fmol per protein) was loaded onto a 300 um i.d. x 15 cm C18 column (LC Packings) and the peptides were separated using a linear acetonitrile/water/TFA gradient (~5 L/min). The LC eluent was automatically deposited onto an unmodifi ed stainless steel MALDI target using the AccuSpot MALDI spotting robot (Shimadzu, Japan). Fractions were collected every 6 sec from T = 3 - 41 min. MALDI matrix solution (CHCA; 5 mg/mL in 50/50 acetonitrile/0.1% TFA) was added and mixed with the LC eluent using the AccuSpot.

iTRAQ sample: 2 samples of an equimolar 5-protein mixture were labelled using the 114 and 117 reagents using the manufacturers recommended procedure. The two samples were combined and prepared for LCMALDI analysis using an SCX trap. 5 L of the digest mixture (500 fmol per protein) was loaded onto a C18 column and prepared as described above.

MALDI-MS and MALDI-MS/MS acquisitions were performed on an AXIMA-TOF2 mass spectrometer (Shimadzu Biotech/Kratos, UK). All acquisitions were performed in fully automated mode using the LC-MALDI software functionality of the MALDI-MS software. The minimum precursor intensity threshold was set to 30 mV and there was no limit on the number of candidates selected for MS/MS (i.e. all precursors meeting the acceptance criteria were selected). For MS/MS acquisitions, helium was used as the collision gas.


Results



Discussion
Protein Mixture Sample

Using the LC-MALDI technique described, we successfully identifi ed 33 p
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. A Positive Selection Assay for Mutation Analysis in Big Blue Animals
2. Epitope-Tagging Vectors for Functional Analysis in Yeast
3. Map and Link Human Genetic Disorders with SSLP Analysis
4. CastAway Precast Gels for Rapid Automated DNA Sequence Analysis
5. Single-cell RT-PCR Analysis of Paramecium Primaurelia with the Eppendorf cMaster RTplusPCR System
6. Structural Analysis of Glycosylated Peptides in Complex Mixtures with Ion Trap MSn
7. Use of Transcriptor Reverse Transcriptase in Microarray Analysis
8. Increased Sensitivity in Microarray Analysis
9. Gene-Expression Analysis of TP, DPD, and TS Using the LightCycler mRNA Quantification KitsPLUS
10. Agilent 2100 Bioanalyzer Automated Analysis System
11. Analysis of Microarray Data
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: MALDI and MALDI for the Analysis Complex Peptide Mixtures

(Date:7/25/2014)... BEIJING , July 25, 2014 Sinovac Biotech ... biopharmaceutical products in China , today announced ... second quarter ended June 30, 2014, after market close on ... conference call prior to the market opening on Friday, August ... at 8:00 p.m. China Standard Time) to review the Company,s ...
(Date:7/24/2014)... Calif. , July 24, 2014  Asterias ... in the emerging field of regenerative medicine, announced ... of a presentation to investors on Tuesday, July ... PDT.  The presentation will include an overview of ... access the webcast, visit http://www.ustream.tv/channel/asterias-biotherapeutics at ...
(Date:7/24/2014)... Distinguished Professor of Engineering at the University of ... solve problems in energy engineering, environmental sustainability and ... zeolites, porous rock with a well-defined, crystalline structure. ... so precisely decided that zeolites can separate molecules ... an angstrom (one-tenth of a nanometer), making them ...
(Date:7/24/2014)... , July 24, 2014 SRI International ... with the National Institute of Allergy and Infectious Diseases ... conduct preclinical development of potential therapies for HIV infection ... to treat HIV and AIDS and the complications and ... microbicides for preventing sexual transmission of HIV. ...
Breaking Biology Technology:Sinovac to Host Conference Call to Report Second Quarter 2014 Unaudited Financial Results 2Asterias Biotherapeutics Announces Live Investor Webcast 2Asterias Biotherapeutics Announces Live Investor Webcast 3Asterias Biotherapeutics Announces Live Investor Webcast 4University of Delaware researcher describes new approach for creating organic zeolites 2National Institute of Allergy and Infectious Diseases Awards SRI International Contract to Study New Therapies for HIV and AIDS 2National Institute of Allergy and Infectious Diseases Awards SRI International Contract to Study New Therapies for HIV and AIDS 3
... SANTA MONICA, Calif., May 19 Cord Blood America, Inc. ... preservation company ( http://www.cordblood-america.com ) focused ... families nationwide and internationally, announced today that it will hold ... Eastern time to discuss 2009 first quarter results for the ...
... Million in New Projects Approved in Atlanta ATLANTA, ... bioscience and biotechnology convention, the Kansas Bioscience Authority ... investments as part of a $581 million initiative ... state.The funding decisions made at the Bio International ...
... -- Prototype of the Innovative Combined Technology on ... 19 Hospira, Inc. (NYSE: HSP ... delivery company, today announced the coming integration of ... EndoTool(R) glucose management software. A prototype of this ...
Cached Biology Technology:Cord Blood America to Hold Conference Call on Thursday May 21st at 11 a.m. Eastern Time to Discuss 2009 First Quarter Results and Corporate Developments 2As Other States Pull Back, Kansas Accelerates Innovation Investments at Bio International Convention 2As Other States Pull Back, Kansas Accelerates Innovation Investments at Bio International Convention 3As Other States Pull Back, Kansas Accelerates Innovation Investments at Bio International Convention 4Hospira Announces Plans to Integrate Symbiq(R) and EndoTool(R) Technologies 2Hospira Announces Plans to Integrate Symbiq(R) and EndoTool(R) Technologies 3
(Date:7/25/2014)... time, Spanish researchers have detected an unknown interaction between ... are introduced into a droplet of salt water and ... crystallisation to create biomineralogical biosaline 3D morphologically complex formations, ... bacteria are revived. The discovery was made by chance ... of the Astrobiology journal and may help ...
(Date:7/24/2014)... New York, NY - July 24, 2014 -- Keryx ... results from the long-term, randomized, active control Phase 3 ... ferric iron-based phosphate binder, for the treatment of hyperphosphatemia ... The PERFECTED study (PhosphatE binding and iRon delivery with ... the Journal of the American Society of Nephrology ...
(Date:7/24/2014)... on average over a 35 year period in which ... impact of humans on declining animal numbers. This decline ... insects, spiders, crustaceans, slugs and worms bring to our ... decomposition for nutrient cycling, water filtration and human health. ... led by UCL, Stanford and UCSB, focused on the ...
Breaking Biology News(10 mins):Bacteria manipulate salt to build shelters to hibernate 2Zerenex™ (ferric citrate) long-term Phase 3 study results published in JASN 2Zerenex™ (ferric citrate) long-term Phase 3 study results published in JASN 3Zerenex™ (ferric citrate) long-term Phase 3 study results published in JASN 4Invertebrate numbers nearly halve as human population doubles 2Invertebrate numbers nearly halve as human population doubles 3
... A gynecologist and a molecular biologist at UT Southwestern Medical Center ... prolapse a condition in which the uterus, bladder or vagina ... a loss of elasticity and a breakdown of proteins in the ... than 50 years of age. Besides creating pelvic pressure, prolapse can ...
... Scientists have long known that people who have a ... of starting to abuse alcohol. The reason for this has ... of Gothenburg, Sweden, provides part of the answer. The study ... an alcoholic react more positively to alcohol than other people. ...
... be published online this week in the Proceedings of ... of a novel hepatitis C-like virus in dogs. The ... into how hepatitis C in humans may have evolved and ... study how it causes disease. The research was ...
Cached Biology News:UT Southwestern researchers find protein breakdown contributes to pelvic organ prolapse 2Heredity behind subjective effects of alcohol 2Discovery of canine hepatitis C virus opens up new doors for research on deadly human pathogen 2Discovery of canine hepatitis C virus opens up new doors for research on deadly human pathogen 3