This core will provide """"""""state of the art"""""""" proteomic and mass spectrometry support for all projects (J. Zheng, H. Gendelman, and Y. Persidsky) in this proposal. This will include protein profiling for neuronal progenitor cells, monocyte-derived macrophages, microglia as well as cell-cell interactions relevant to studies that affect virus and cell-protein changes in brain mononuclear cell inflammatory responses and for developmental therapeutics in NeuroAIDS. We will use 2-dimensional electrophoresis (2DE) with Difference Gel Electrophoresis (DICE) (2DE DICE) as a primary profiling approach. To complement these established methods, we will use an ABI4800 MALDI-TOF/TOF (Applied Biosystems, Inc.) for rapid and precise mass analysis. Post-translational modifications such as phospnorylation will be done with a QQ-LIT (QTrap4000, Applied Biosystems, Inc.). In addition, we have quantitative mass spectrometry systems that incorporates isotope-coded affinity tags (ICAT), isotope Tags for Relative and Absolute Quantification (iTRAQ)?, and Stable Isotope-labeling with Amino Acids in Cell culture (SILAC) for protein labeling or tagging. Our ongoing collaborations with Beckman-Coulter, Inc. have extended our protein capabilities with the acquisition of a Proteomel_ab?PF2D Protein Fractionation System for the profiling complex protein samples. Taken together, this core has evolved from simple surface enhanced laser desorption ionization assays included in the initial program project proposal to become a university-wide well-integrated core facility, which serves to greatly facilitate all the proposed research included in the application.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Program Projects (P01)
Project #
5P01NS043985-07
Application #
7983554
Study Section
National Institute of Neurological Disorders and Stroke Initial Review Group (NSD)
Project Start
Project End
Budget Start
2009-03-01
Budget End
2010-02-28
Support Year
7
Fiscal Year
2009
Total Cost
$126,784
Indirect Cost
Name
University of Nebraska Medical Center
Department
Type
DUNS #
168559177
City
Omaha
State
NE
Country
United States
Zip Code
68198
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