Navigation Links
Direct On-Membrane MS/MS Analysis using the Chemical Printer (CHIP-1000) and AXIMA-QIT


Microscale on-membrane PMF analysis can be conducted on proteins transferred to a PVDF membrane utilizing the microdispensing function of a chemical printer (Refer to AXIMA Application 14, 17). Moreover, using the AXIMA-QIT ion trap MS in conjunction with the chemical printer enables MS analysis with high mass accuracy (<0.1Da). In this application, we succeeded in conducting MS/MS analysis of proteins immobilized on PVDF transfer membranes, something which has been difficult up to now due to the irregularities of the membrane surface and electrical charge. Direct on-membrane MS/MS analysis using the chemical printer together with the AXIMA-QIT, in addition to the established methodology for onmembrane PMF analysis, is expected to be extremely effective in identification of unknown proteins.

Protein spots from E. coli separated by 2-dimensional electrophoresis were transferred to a PVDF membrane for use as the sample (Fig. 1). A chemical printer was then used to perform enzymatic digestion (trypsin digestion) on selected protein spots. Following enzymatic digestion, matrix solution was microdispensed on each print position, and onmembrane MS analysis was then conducted using the AXIMA-QIT. The obtained MS spectra are shown in Fig. 2.




Next, the AXIMA-QIT was used to conduct on-membrane MS/MS analysis of precursor ions identified from characteristic signals of protein spots verified through PMF analysis. We were able to identify the respective protein spots by conducting an MS/MS ion search based on the obtained MS/MS spectra.

Fig. 1 shows a portion of the transfer membrane stained with Direct Blue 71 after transfering E. Coli extract separated by 2-dimensional electrophoresis to the PVDF membrane. The MS spectra obtained from microscale on-membrane PMF analysis using the chemical printer are shown i
'"/>

Source:


Page: All 1 2

Related biology technology :

1. Counting Colonies or Plaques Directly from Images of Agar Plates
2. High-Efficiency Site-Directed Mutagenesis of Large Plasmid Vectors
3. Constructing Directional cDNA Libraries from Limited Amounts of RNA
4. Generate High-Quality, Directional Plasmid cDNA Libraries
5. Optimizing the Direct Amplification of Missing cDNA 5 Ends Using the Eppendorf Mastercycler gradient
6. Optimizing the Direct Amplification of Missing cDNA 5 Ends Using the Eppendorf Mastercycler gradient
7. TripleMaster PCR System: Direct Comparison with Competitors
8. DirectPrep 96 Miniprep System for cost-effective, high-throughput plasmid DNA purification
9. A Further Step in Understanding Apoptosis Direct Detection of PARP Cleavage
10. YOUR DATA: Direct Real-time RT-PCR on Sympathetic Neuron Lysates Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs
11. YOUR DATA: Direct Real-time RT-PCR on Sympathetic Neuron Lysates Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Direct Membrane Analysis using the Chemical Printer CHIP and AXIMA QIT

(Date:8/21/2014)... August 21, 2014 2014 ... Industry” is a professional and in-depth research ... report provides basic Seaweed Fertilizer information, including ... chain structure as well as industry overview. ... including domestic market as well as global ...
(Date:8/21/2014)... New Rochelle, N.Y. (PRWEB) August 21, 2014 ... it is the first private college in the Mid-Hudson ... The College will now begin accepting applications from qualified ... , “We are very pleased to have been ... to support Governor Cuomo’s transformative initiative to stimulate economic ...
(Date:8/21/2014)... , Aug. 21, 2014  China Cord Blood Corporation (NYSE: ... plan to release financial results for the first quarter ... after market close in the US.  ... a.m. ET on Friday, August 29, 2014 to discuss ... the Company,s recent developments, followed by a question and ...
(Date:8/21/2014)... St. Louis, MO (PRWEB) August 21, 2014 ... premier, full service biopharmaceutical contract development and manufacturing ... selected by Progenics Pharmaceuticals, Inc., an oncology company ... targeting and treating cancer, to manufacture the anti-prostate ... Progenics’ PSMA ADC product candidate. Under the ...
Breaking Biology Technology:Internationals Seaweed Fertilizer Industry 2014 Development Trend Analysis in New Research Study at DeepResearchReports.com 2Internationals Seaweed Fertilizer Industry 2014 Development Trend Analysis in New Research Study at DeepResearchReports.com 3The College of New Rochelle Becomes First Private College in Mid-Hudson Region to Earn START-UP NY Designation 2The College of New Rochelle Becomes First Private College in Mid-Hudson Region to Earn START-UP NY Designation 3China Cord Blood Corporation to Report First Quarter of Fiscal 2015 Financial Results 2Gallus Enters Agreement with Progenics Pharmaceuticals, Inc. for Manufacture of Anti-PSMA Monoclonal Antibody 2Gallus Enters Agreement with Progenics Pharmaceuticals, Inc. for Manufacture of Anti-PSMA Monoclonal Antibody 3
... strategies to create vaccines for avian flu through ... genetically-engineered virus-like particles (VLPs), ROCKVILLE, Md., Nov. 13 ... its strategy for treating avian flu,through genetically-engineered virus-like particles ... Clinic 6th Annual Medical Innovation,Summit., Novavax, Inc. believes ...
... Genaera,Corporation (Nasdaq: GENR ) will present at ... Center in Mannheim, Germany. Dr. Henry,Wolfe, Executive Vice ... overview on Tuesday, November 18, 2008 at 11:00 ... is developing trodusquemine (MSI-1436) for type 2,diabetes and ...
... Covanta Holding,Corporation (NYSE: CVA ), a world ... renewable energy facilities, today,announced that it has amended ... co-owners Ridgewood Maine, L.L.C. and Indeck Energy,Services, Inc., ... agreed to acquire these,two facilities for approximately $52 ...
Cached Biology Technology:Cleveland Clinic Recognizes New Strategies for Creating Vaccines for Avian Flu as a Top Ten Medical Innovation for 2009 2Cleveland Clinic Recognizes New Strategies for Creating Vaccines for Avian Flu as a Top Ten Medical Innovation for 2009 3Cleveland Clinic Recognizes New Strategies for Creating Vaccines for Avian Flu as a Top Ten Medical Innovation for 2009 4Genaera to Present at BIO-Europe 2008 2Covanta Announces Amended Terms for its Acquisition of Two Biomass Energy Facilities in Maine 2Covanta Announces Amended Terms for its Acquisition of Two Biomass Energy Facilities in Maine 3Covanta Announces Amended Terms for its Acquisition of Two Biomass Energy Facilities in Maine 4
(Date:8/21/2014)... BATON ROUGE In a finding that has implications ... and planets elsewhere in the solar system, LSU Associate ... funded by the National Science Foundation, or NSF, this ... sediments of a lake that lies 800 meters (2600 ... sheet support "viable microbial ecosystems.", Given that more than ...
(Date:8/21/2014)... A study recently published in the journal Carcinogenesis ... a new role for the protein adenomatous polyposis coli ... of cancer-related deaths in the U.S. , Lead author ... Biosciences and co-leader of the Cancer Biology program at ... of her career trying to understand the various activities ...
(Date:8/21/2014)... State University have developed a novel and versatile modeling ... for creating new materials as well as for studying ... allows us to model much larger and more complex ... says Nan Li, lead author of a paper on ... Department of Materials Science and Engineering. "This is a ...
Breaking Biology News(10 mins):800 meters beneath Antarctic ice sheet, subglacial lake holds viable microbial ecosystems 2800 meters beneath Antarctic ice sheet, subglacial lake holds viable microbial ecosystems 3800 meters beneath Antarctic ice sheet, subglacial lake holds viable microbial ecosystems 4Research offers insight into cellular biology of colorectal cancer 2Research offers insight into cellular biology of colorectal cancer 3Researchers develop models to study polyelectrolytes, including DNA and RNA 2
... Cleveland Clinic and Riverain Medical today announced the ... to determine whether chest X-ray CAD (computer-aided detection) can ... program is designed to determine whether chest X-ray CAD ... an early stage when they are most treatable, leading ...
... arrived at the Red Planet. Almost four years later, ... its instruments send back a stream of images and ... its remarkable career by completing 5000 orbits of Mars. ... orbiter has revolutionised our knowledge of Mars, probing every ...
... human immunodeficiency virus (HIV-1) molecular machinery responsible for ... a study by investigators at the University of ... published November 23 in PLoS Computational Biology. ... of HIV-1 has so far foiled the best ...
Cached Biology News:Cleveland Clinic leading clinical program to improve early-stage lung cancer detection 2Mars Express -- 5000 orbits and counting 2Mars Express -- 5000 orbits and counting 3
... Use in any standard fixed-angle ... 16,000 g.,Spin- centrifuge tube filters ... within a centrifuge tube. They filter ... removal, HPLC sample preparation, removal of ...
... treated (or medium binding) polystyrene surface ... biomolecules through passive interactions,• Is suitable ... molecules, such as antibodies, that have ... with the surface,• Binding capacity of ...
... depth • Reusable, press-to-seal gasket and cover ... and disassembled for hybridizing and imaging thick ... with up to a 13 mm diameter. ... Minimize specimen handling, eliminate evaporation and leaking ...
Comp Dim: chamber diam. depth 20 2.8 mm chamber maximum volume 720 μL...
Biology Products: