Navigation Links
Detection, identification, and quantitation of an olive allergen using Ettan MDLC, MS/MS, and DeCyder MS

LC grade


Methods
Protein samples
Protein samples were obtained from the pollen of the olive tree, Olea europaea, by aqueous extraction at low pH. The two samples used in this study, Ole_e1_"mer" and Ole_e1_"lite," refer to different batches of collected pollen containing varying amounts of the allergen.

Protein fractionation
Two mg each of Ole_e1_"mer" and Ole_e1_"lite" was partially purified using an Ettan LC System. The chromatographic separation was performed on a Superdex 75 10/300 GL gel filtration column at 0.5 ml/min in 80 mM sodium acetate, pH 4.5, 50 mM NaCl, and 0.5-ml fractions were collected using Fraction Collector Frac-950. Fractions were examined using MALDI-ToF MS. Those containing proteins with molecular weights 14–20 kDa were pooled and dried in vacuo.

Protein reduction, alkylation, and trypsin digestion
The pooled, dried fractions from the gel filtration step were dissolved in 0.5 ml of 9 M urea containing 50 mM DTT and incubated at ambient temperature for 60 min. An equal volume of a solution consisting of 8 M urea, 250 mM Tris-HCl, pH 8.8, and 125 mM iodoacetamide was added, and incubation was continued for another 60 min in darkness. The samples were desalted against 20 mM ammonium bicarbonate on NAP-10 Columns, and then digested with 5 µg trypsin for 2 h at 37 °C. The digestions were stopped by acidification with formic acid.

Dual reversed-phase nanoscale liquid chromatography
Ettan MDLC was used in the high-throughput configuration using two reversed-phase (RPC) trap columns 300 µm i.d. x 5 mm (Zorbax 300-SB C18) for online desalting and sample cleanup, followed by two nanoscale RPC analytical columns 75 µm i.d. x 150 mm (Zorbax 300-SB C18) for high-resolution separation. One set of RPC trap/analytical columns was equilibrated while the second set
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9

Related biology technology :

1. Comparison of different protein quantitation methods
2. Detection and quantitation of low-abundant proteins with ECL Plex fluorescent Western blotting
3. High sensitivity quantitation of metabolites of nitrofuran antibiotics in animal tissue using LC/MS/MS
4. Outstanding transfection rates using the Multiporator
5. Outstanding transfection rates using the Multiporator
6. Isolation of Cosmids using Eppendorfs Perfectprep Plasmid Mini Kit
7. Normalizing cDNA libraries using the EppendorfThermomixer
8. Optimizing DNA Amplification Protocols using the Eppendorf Mastercycler
9. Efficient DNA transfection of primary CNS neurons using TransMessenger Transfection Reagent
10. Improved Quantitative Selectivity of Clenbuterol in Human Urine using High Resolution on the TSQ Quantum Mass Spectrometer
11. Efficient RNAi-mediated gene silencing in neuronal cells using QIAGEN siRNA and TransMessenger Transfection Reagent*
Post Your Comments:
(Date:1/15/2014)... Village, Calif. (PRWEB) January 15, 2014 ... treatment and management through the genomic analysis of tumor ... offer Therapy Finder™, a web-based cancer decision support application ... reporting option with Cynvenio’s ClearID™ genomic test, Therapy Finder ...
(Date:1/15/2014)... Houston, TX (PRWEB) January 15, 2014 ... Alzheimer’s disease, and about 1 in 3 seniors will ... ( source ). These jaw-dropping figures have shocked many ... function and, hopefully, help prevent these tragic age-related cognitive ...
(Date:1/15/2014)... CA (PRWEB) January 15, 2014 ... research solutions, today announced that Lupin Limited, one of ... Freeslate’s CM Protégé PharmD System for high ... Mumbai, India, is focused on a wide range of ...
(Date:1/14/2014)... 2014 Histogen, Inc., a regenerative medicine ... grown under simulated embryonic conditions, today announced that they ... Medical, Inc. for physician-dispensed aesthetic products containing Histogen’s proprietary ... agreement is an amendment to the existing license between ...
Breaking Biology Technology:Cynvenio Offers New Breast Cancer Therapy Finder Application for its ClearID Breast Cancer Genomic Test 2Cynvenio Offers New Breast Cancer Therapy Finder Application for its ClearID Breast Cancer Genomic Test 3Biohack Pure Offers 5 Tips for Increasing Memory in 2014 2Lupin Selects Freeslate’s CM Protégé PharmD System to Accelerate Polymorph Screening for Drug Development 2Lupin Selects Freeslate’s CM Protégé PharmD System to Accelerate Polymorph Screening for Drug Development 3Histogen and Suneva Medical Expand License for Cell Conditioned Media-based Aesthetic Products Internationally 2Histogen and Suneva Medical Expand License for Cell Conditioned Media-based Aesthetic Products Internationally 3
... April 24, 2008 / b3c newswire / - ... are working on a project to synthesize a gene ... statistically expected and controlled ratio at preselected positions within ... core technology, Slonomics™, a unique DNA engineering platform using ...
... N.C. -- Duke University chemists have found a way ... thick in very large numbers, removing a major roadblock ... These single-walled carbon nanotubes also follow parallel paths as ... potentially impede electronic performance, said Duke associate chemistry professor ...
... Sangamo,BioSciences, Inc. (Nasdaq: SGMO ) today announced ... results on Wednesday, April 30,2008, after the market closes. ... at 2:00 p.m. PT, which will be open to ... the company will review,the financial results and discuss other ...
Cached Biology Technology:Sloning and the Kornberg group at Stanford University starting evaluation project regarding the synthesis of a Statistically Randomized Gene Library 2Nanotubes grown straight in large numbers 2Nanotubes grown straight in large numbers 3Sangamo BioSciences Announces First Quarter Conference Call and Webcast 2Sangamo BioSciences Announces First Quarter Conference Call and Webcast 3
(Date:4/22/2014)... Analyzing thousands of breeding bird surveys sent in ... Canada over 35 years, wildlife researchers report that most ... northward or toward higher elevation in response to climate ... means that most previous studies of potential climate change ... or the other have likely underestimated the effects of ...
(Date:4/22/2014)... effects of temperature and precipitation on the global distribution ... of climate research, the UV-B radiation, is often neglected. ... colleagues from the Universities in Olomouc (Czechia), Halle and ... space agency in such a way that they can ... on organisms. , The basic input data were provided ...
(Date:4/22/2014)... your age" is another way of asking him or ... bring improvements. Certain cells of the immune system tend ... to illness. Because these cells are known to misbehave ... microgravity on immune cells to better understand how our ... the National Institute on Aging, part of the National ...
Breaking Biology News(10 mins):Wildlife response to climate change is likely underestimated, experts warn 2UV-radiation data to help ecological research 2For an immune cell, microgravity mimics aging 2For an immune cell, microgravity mimics aging 3For an immune cell, microgravity mimics aging 4For an immune cell, microgravity mimics aging 5
... , WEST LAFAYETTE, Ind. - Scientists could take greater ... of advanced techniques used to understand the mechanisms that ... former Purdue University researchers say. In a perspective ... Purdue assistant professor of genetics, and Ivan Baxter, a ...
... 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 ...
... big boss proteins that switch on broad developmental ... regulators of muscle development or fat metabolism. One ... protein, takes charge following a cellular crisis known ... by the accumulation of misfolded and aggregated proteins. ...
Cached Biology News:Adoption of advanced techniques could propel crop improvement 2Discovery may lead to new tomato varieties with vintage flavor and quality 2Forty's a crowd 2Forty's a crowd 3Forty's a crowd 4
Immunochemistry 1 & 2...
...
Hucal Agx Antigen Expression Service Specificity: Hucal Agx ...
MHC. Sections include: the isolation and functional properties of antigen-presenting cells analysis of MHC class II endocytosis and peptide turnover analysis of signalling via MHC class II molecu...
Biology Products: