Purdue University and the University of Alabama at Birmingham
Botanicals Center for Age-Related Disease
| Monday, September 11 |
| 8:00 - 8:30am |
Breakfast |
| 8:30 - 8:45am |
Introduction to Workshop - S. Barnes/H. Kim/JM. Wyss |
| 8:45 - 9:15am |
Selection of animal models - D. Waters (Purdue)
- Animals: knockout, transgenic; Cells (mechanism) |
| 9:15 - 10:00 am |
Collecting biological samples - H. Kim, S.
Wang, JM. Wyss
- metabolic cages, blood sampling, interstitial fluids/aqueous humor, frozen tissues, subcellular fractionation |
| 10:00 - 10:20 am |
Coffee break |
| 10:20 - 11:00 am |
Selection of botanicals - S. Barnes
- previous use by other investigators, defined product, safety data, dose selection, existing pharmacokinetic data, dealing with the supplier/manufacturer, mixing botanicals in the diet, sampling the diet |
| 11:00 - 11:30 am |
Analysis of the botanical - J. Prasain
- extraction and LC methods (UV diode, electrochemical)
|
| 11:30 - 12 noon |
LC-MS methods - S. Barnes, J. Prasain
- Ionization methods (ESI, HN-APCI), Qualitative analysis (MS, MS-MS), Quantitative analysis (SIM and MRM) |
| 12:00 - 1:00 pm |
Lunch |
| 1:00 - 1:40 pm |
Characterizing the animal model - JM. Wyss
- blood pressure, maze, cataracts |
| 1:40 - 2:20 pm |
Physical methods in models of cataract disease - O. Srivastava |
| 2:20 - 3:00 pm |
Studying botanicals and skin models of disease - S. Katiyar |
| 3:00 - 3:20 pm |
Coffee break |
| 3:20 - 5:00 pm |
The Botanical Center Facilities |
| Tuesday, September 12 |
| 8:00 - 8:30 am |
Breakfast |
| 8:30 - 8:35 am |
Introductions (for proteomics workshop guests) |
| 8:35 - 8:55 am |
Biology: the basis for smart proteomic approaches to protein analysis - H. Kim
- Choice of organ or cells, whole cell lysate, subcellular fractionation, tagged complexes |
| 8:55 - 9:15 am |
Proteomics and Thiol Modification - A. Landar
- 2D-gels (IEF/SDS, blue native) |
| 9:15 - 9:35 am |
Protein purification prior to proteomics analysis - S. Barnes
- Chromatofocusing, Anion/cation exchange,
Hydrophobic, Gel filtration |
| 9:35 - 9:55 am |
Detecting and quantifying proteins on gels - H. Kim
- Staining methods (Coomassie blue, Sypro ruby, Pro-Q diamond, etc.)
- Cy-Dye labeling |
| 9:55- 10:15 am |
Statistical Experimental Design in Proteomics - S. Meleth
- DeCyder
|
| 10:15 - 10:40 am |
Coffee break |
| 10:40 - 11:10 am |
Phosphorylationand Glycosylationby ECD - M. Renfrow
- MALDI, electrospray ionization
- Detectors (TOF and TOF/TOF, Quadrupole, FT-ICR, Orbitrap, Hybrid detectors)
- mass accuracy and mass resolution |
| 11:10 - 11:40 am |
Qualitative Proteomics - J. Mobley
- peptide mass fingerprinting (MASCOT), peptide MUDPIT (2D-SAX-rpLC-tandem mass spectrometry) (SEQUEST) |
| 11:40 - 12:10 pm |
Quantification in the world of proteomics - S. Barnes
- peptide coverage, iCAT labeling, iTRAP labeling, 180/160 labeling |
| 12:15 - 1:15 pm |
Lunch |
| 1:15 - 2:15 pm |
Detection of post-translational modifications of peptides - S.
Barnes
Post-translational Modifications to Human Bile Acid CoA:Amino Acid Nacyltransferase - E. Shonsey
Localization of 4HNE modifications - S. Eliuk |
| 2:15 - 3:00 pm |
Profiling and Imaging Mass Spectrometry - J. Mobley |
| 3:00 - 3:20 pm |
Coffee break |
| 3:20 - 5:00 pm |
Demonstrations of proteomics and mass spectrometry applications at UAB |
