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:4/23/2015)... 23, 2015  Propanc Health Group Corporation (OTCBB: ... on the development of cancer treatments for patients ... product includes PRP, a patented formulation consisting of ... well as the enzyme amylase designed to synergistically ... combination of anti-cancer agents working in combination with ...
(Date:4/23/2015)... FRANCISCO, CA (PRWEB) April 23, 2015 ... delivery, today announced that the United States Patent and ... the Company’s NANOPOR™ technology. US Pat. ... covers Delpor’s microfabricated drug delivery device for ... Delpor’s proprietary NANOPOR™ technology for the sustained (zero-order) release ...
(Date:4/22/2015)... April 22, 2015 /PRNewswire/ - BioAmber Inc. (NYSE and EURONEXT Paris:BIOA) today announced ... quarter 2015 on Thursday, May 7, 2015 after the ... webcast and conference call for 4.30 pm ET (1.30 ... Live audio of the conference call will be webcast ... members of the news media and the general public. ...
(Date:4/22/2015)... , April 22, 2015 Steep ... analytics, announced today that it has opened an ... continued growth and expansion.  The common stock investment ... Section 506(c), a law intended to encourage funding ... Prior to 506(c), small businesses largely relied on ...
Breaking Biology Technology:Early Stage BioTech offers Plenty of Risk; Propanc Health Group Analyst Brief Issued by BrokerBank Securities, Inc. 2Delpor, Inc. Announces the Issuance of 2 New US Patents Based on its Nanopore Technology which Enables the Sustained Release of Drugs from a Small Implant Device 2Delpor, Inc. Announces the Issuance of 2 New US Patents Based on its Nanopore Technology which Enables the Sustained Release of Drugs from a Small Implant Device 3BioAmber Announces Webcast of Fiscal First Quarter 2015 Financial Results 2Steep Hill Launches $5M Limited Stock Offering 2Steep Hill Launches $5M Limited Stock Offering 3
... Amex: NBS ) ("NeoStem" or the "Company"), an ... today announced that the Company and its wholly-owned subsidiary, Progenitor ... conferences: the 2011 World Stem Cell Summit, the AABB Annual ... 2011 World Stem Cell Summit – ...
... Oct. 4, 2011 Exosome Diagnostics, a leading ... in personalized medicine, today announced the appointment of ... Donovan, M.D., Ph.D., as chief medical officer, and ...  Raphael Bueno, M.D., associate chief of cardiothoracic surgery ...
... have discovered two early-stage phases of carbon nanotube growth ... tangle of tube growth that ultimately yields to orderly ... in the latest edition of the journal Nanotechnology ... Professor of Physics Zhifeng Ren and researcher Dr. Hengzhi ...
Cached Biology Technology:NeoStem Announces Participation in Multiple October Conferences 2NeoStem Announces Participation in Multiple October Conferences 3NeoStem Announces Participation in Multiple October Conferences 4Exosome Diagnostics Announces Additions to Management and Scientific Advisory Board as Company Expands Clinical Validation Studies in Oncology 2Exosome Diagnostics Announces Additions to Management and Scientific Advisory Board as Company Expands Clinical Validation Studies in Oncology 3Exosome Diagnostics Announces Additions to Management and Scientific Advisory Board as Company Expands Clinical Validation Studies in Oncology 4Exosome Diagnostics Announces Additions to Management and Scientific Advisory Board as Company Expands Clinical Validation Studies in Oncology 5Boston College Researchers discover 2 early stages of carbon nanotube growth 2
(Date:3/31/2015)... , March 31, 2015   Guidepoint , a ... of the Post-Surgical Pain Management TRACKER, part of the ... usage trends in the medical device and therapeutics markets. ... data including treatment volumes, market share, and adoption rates ... associated with joint surgery. The Post-Surgical Pain ...
(Date:3/24/2015)... DUBLIN , Mar. 24, 2015 Research and ... addition of the "Global Iris Recognition Market 2015-2019" ... forecast the Global Iris Recognition market to grow at ... This report covers the present scenario and ... for the period 2015-2019. To calculate the market size, ...
(Date:3/24/2015)... PALM BEACH GARDENS, Fla. , March 24, ... of biometric identity management solutions, today announced that ... enabling premium clothing accessory retailer Goorin Bros ... login. The Crossmatch biometrics-based solution secures access to ... customers from the increased risk of data breaches. ...
Breaking Biology News(10 mins):Guidepoint Launches Post-Surgical Pain Management TRACKER 2Guidepoint Launches Post-Surgical Pain Management TRACKER 3Global Iris Recognition Market 2015-2019 with 3M Cogent, Bioenable Technologies, Cross Match Technology, Iris ID Systems & Safran Dominating 2Crossmatch Fingerprint Readers Help Retailer Prevent Data Breaches at Point-of-Sale 2
... Cancer Cell describes a small molecule inhibitor of polo-like kinase1 ... cancer therapy. The compound, ON01910, is a potent inhibitor of ... toxicity. The results of this study have led to clinical ... Plk1 is a molecule that plays a key role in ...
... proteins called phosphatases play a key role in the ... fruit flies and mice by researchers at the University ... collaboration with scientists at the Salk Institute, La Jolla, ... stem cells and for silencing expression of neuronal genes ...
... toward understanding the regulation of blood stem cells and ... research study published in the February issue of Developmental ... to examine the role of a key gene known ... from this research will be useful for future studies ...
Cached Biology News:New drug shows promise as powerful anticancer agent 2New insight into regulation of blood stem cells 2
BD BioCoat Collagen IV 150 mm Culture Dishes...
Polymerase (RNA) II (DNA directed) polypeptide L...
Chicken Anti-Human DNA-directed RNA Polymerase II 7.6 Kd Polypeptide (POLR2L Polyclonal AntibodyOther Sub-Family: not assigned-Other Applications: ELISA, Western Blot...
The BD CBA Human Apoptosis Kit can be used to quantitatively measure,cleaved PARP, Bcl-2, and active Caspase-3 protein levels in a single sample. The,Kit performance has been optimized for analysis o...
Biology Products: