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

cting the fractions, and then injecting these onto a nanoscale LC, the peptides of low abundance, such as phosphopeptides, could be detected (3). Forty micrograms of a mouse brain tissue tryptic digest was first separated by SCX using salt gradient elution. The chromatogram is shown in Figure 2.

Thirty of the collected fractions were further analyzed by LC-MS using the neutral loss function. Phosphopeptides were found in one-third of the analyzed SCX fractions as shown in Figure 2. They mainly eluted early in the salt gradient due to their reduced charge state (4). SCX can therefore be used as a phosphopeptide enrichment strategy.

An ion chromatogram and the MS3 events (i.e. where a neutral loss was detected) from one of the fractions are shown in Figure 3.

The phosphopeptides were found and the phosphorylation sites identified by TurboSEQUEST database searches on all MS3 spectra. The results were confirmed manually by studying the raw spectra. It was important to confirm that the charge state of the peptide was correct, that the neutral loss ion dominated the MS/MS spectrum (Fig 4), and that the sequence data was of high quality.

Database searches were then performed on all MS/MS spectra, and the results were used to confirm the MS/MS searches and to find tyrosine phosphorylations. Phospho-tyrosine does not lose phosphoric acid during collision in the ion trap; therefore, sequence data from MS/MS was used to find these phosphorylations (+80). Some possible tyrosine phosphorylations were identified but need further evaluation. Tyrosine-phosphorylated proteins exist in very low abundance in cells and probably fall below the detection limit of the method.

In total, 60 phosphorylated peptides were found originating from 50 proteins. Some of the identified phosphorylation sites are shown in Table 1. The proteins presented in the table are al
'"/>

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:1/15/2014)... TaiGen Biotechnology Company, Limited ("TaiGen") today announced that they ... Russian pharmaceutical company, to develop and commercialize nemonoxacin (Taigexyn ... , Turkey and other members ... antibiotic for the treatment of bacterial infections including those ...
(Date:1/14/2014)... , Jan. 14, 2014  3D Communications, a leading provider of strategic ... business, and media events in the United States ... associate Virginia Cox , JD, is returning to the firm,s ... Cox re-joins 3D after more than two years of service ...
(Date:1/14/2014)... BETHESDA, Md. , Jan. 14, 2014  Organovo ... institutes from the National Institutes of Health (NIH) to ... more effective treatments to patients on a faster timeline. ... Translational Sciences (NCATS) and the National Eye Institute (NEI) ...
(Date:1/14/2014)... 14, 2014 Global Record Systems, ... information technology solutions for patients, physicians, the biopharmaceutical ... announced today the signing of a three-year Research ... Drug Administration (FDA). This initiative is designed ...
Breaking Biology Technology:TaiGen Biotechnology Signed Exclusive License Agreement with R-Pharm for Nemonoxacin (Taigexyn(R)) 2TaiGen Biotechnology Signed Exclusive License Agreement with R-Pharm for Nemonoxacin (Taigexyn(R)) 3TaiGen Biotechnology Signed Exclusive License Agreement with R-Pharm for Nemonoxacin (Taigexyn(R)) 4Former FDA Associate Commissioner Returns To 3D Communications 2Organovo Announces Collaboration with National Institutes of Health 2Organovo Announces Collaboration with National Institutes of Health 3Global Record Systems Announces Research Collaboration Agreement with FDA to Create a Novel “Big Data” Paradigm for Collection of Patient Safety and Outcomes Information 2
... SHANGHAI, June 29 /PRNewswire-Asia/ -- Sundia MediTech Company, ... Shanghai,China announced today that Dr. Tong Xu has been ... Prior to joining Sundia, Dr. Xu worked ... in San Diego, USA for 12 years. At ...
... Mylan Inc. (Nasdaq: MYL ) today ... with Biocon Limited, a publicly traded company on ... on the development, manufacturing, supply and commercialization of ... global marketplace. Through this partnership, Mylan and Biocon ...
... UPPSALA, Sweden, June 29 , ... Will Fund Gyros to Profitability , ... development, announces that it has raised SEK 80 million,(approximately euro ... Company,s leading shareholder. The new funds will enable Gyros to ...
Cached Biology Technology:Sundia MediTech Appoints Dr. Tong Xu as VP of Process Chemistry 2Mylan Announces Strategic Collaboration with Biocon to Enter the Global Generic Biologics Market 2Mylan Announces Strategic Collaboration with Biocon to Enter the Global Generic Biologics Market 3Mylan Announces Strategic Collaboration with Biocon to Enter the Global Generic Biologics Market 4Gyros Raises SEK 80 Million to Accelerate the Commercialization of its Gyrolab(R) Systems for the Cost-Efficient Development of Biopharmaceuticals 2Gyros Raises SEK 80 Million to Accelerate the Commercialization of its Gyrolab(R) Systems for the Cost-Efficient Development of Biopharmaceuticals 3Gyros Raises SEK 80 Million to Accelerate the Commercialization of its Gyrolab(R) Systems for the Cost-Efficient Development of Biopharmaceuticals 4
(Date:7/9/2014)... Jena (Germany) They are amongst the most numerous inhabitants ... Emiliania huxleyi . Not visible to the naked eye, ... square kilometer sized patches, they are even visible on ... is responsible for approximately half of the global ... Friedrich Schiller University Jena (Germany). In the process the ...
(Date:7/9/2014)... they are used in the right ways. ... Hospital and coauthors analyzed six use cases with ... decompensation (when a patient,s condition worsens); adverse events; ... organ systems. They suspect that cost-savings benefits will ... all six scenarios will be significant. The authors ...
(Date:7/8/2014)... LAKE CITY, July 9, 2014 Collectors found the first ... in west Texas. Then, for two decades, the 14-inch-tall plant ... a third. , Now after a long search turned ... Utah botanist and her colleagues identified the spiny plant as ... heart" in Latin because it was found in Valentine, Texas, ...
Breaking Biology News(10 mins):Short circuit in the food web 2The impact of big data on health care: Health Affairs' July issue 2New plant species from the heart of Texas 2New plant species from the heart of Texas 3New plant species from the heart of Texas 4New plant species from the heart of Texas 5
... Space is not a fun place to get a stomach ... Utah chemists developed a two-minute water quality monitoring method that ... Station. "Now they bring water back on the space ... there is a big delay. You,d like to be able ...
... difference can a tenth of a nanometer make? When it ... resolution of that miniscule amount can mean the difference between ... Case in point: New, improved-resolution views of a zinc transporter ... Laboratory provide not just a structure but also a suggested ...
... stamp that can sniff out poisonous gases or deadly toxins ... 13 issue of the journal Nature Chemistry , Kenneth ... developed an artificial nose for the general detection of toxic ... and works by visualizing odors. This sensor array could ...
Cached Biology News:Water quality in orbit 2Water quality in orbit 3Water quality in orbit 4High-res view of zinc transport protein 2High-res view of zinc transport protein 3Opto-electronic nose sniffs out toxic gases 2Opto-electronic nose sniffs out toxic gases 3
...
The S. cerevisiae Hansen strains are the parental yeast strains from which the deletions were derived....
The DNA DipStick Kit is ideal for estimating nucleic acid concentrations and approximate yields. Works with ss- and ds-DNA, RNA, and oligonucleotides at concentrations as low as 0.1 ng/l....
... The NEW HandyStep electronic from BRAND ... pipetting. It is the only electronic, ... standard syringe tips, including BRAND PD-Tip ... Plus, Fisherbrand Dispenser Tips,and VWRbrand Combi-Syringes. ...
Biology Products: