The following services can be performed by UA Proteomics. Please contact us for further information.
Protein Electrophoresis and Mass Spectrometry Packages:
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2D-PAGE Proteomic Analysis |
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2D-DIGE Proteomic Analysis |
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2D-LC-MS/MS (MudPIT Analysis) |
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Protein Electrophoresis and Purification Services:
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Albumin Depletion of Serum |
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Electrophoresis (1D, 2D, & DIGE) |
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Image Analysis (Typhoon Scanner) |
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Protein Gel Staining (Silver, Coomassie, SPRO Ruby) |
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PhosphoProtein Gel Staining (Pro-Q) |
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Robotic Spot Picking |
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Automated In-Gel Digestion |
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Protein Purification (HPLC, FPLC) |
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Mass Spectrometry Services:
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Mass Spectrometry (nanoLC-MS/MS |
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SEQUEST Database Searching |
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2D-PAGE Proteomic Analysis Package:
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2D-PAGE Gel Electrophoresis |
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Post Staining |
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Simple image analysis |
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In-gel digesion, nanoLC-MS/MS, SEQUEST database searching for up to 12 spots |
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2D-Page with spots selected for processing |
2D-DIGE Proteomic Analysis Package:
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2D-DIGE Gel Electrophoresis on 24cm IPG Strip |
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Scanning on Typhoon |
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Post staining |
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Image analysis with Decyder software to determine gel differences |
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In-gel digesion, nanoLC-MS/MS, SEQUEST database searching for up to 12 spots |
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Multiplexed image received from scanning on the Typhoon |
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Decyder analysis showing similar (green), increased (red), and decreased (blue) spots |
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Decyder analysis showing histogram view of similar (green), increased (red), and decreased (blue) spots. Y axis is the volume of the spot, while X axis is the amount of difference |
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Sample Decyder analysis of a control gel, which is the comparison of the same sample labelled with Cy2 and Cy3. |
Non-gel based method -- LC/LC-MS/MS:
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Protein sample digested with trypsin in solution (caution: cellular lysates containing large amounts of proteases or protease inhibitors are problematic as they interfere with the essential trypsin digestion) |
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Multistep on-line fractionization |
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8-10 cation exchange chromatography steps |
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Each ion exchange step is followed by reverse phase HPLC |
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Provides identification of many (often hundreds) of proteins |
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Albumin Depletion of Serum by SepproTM Column:
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Currently using Genway Pre-packed SepproTM MIXED6 column for removal of the most abundant 6 proteins |
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Affinity based on avian antibody (IgY)-antigen interactions |
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Removes Albumin, IgG, IgA, IgM, Transferrin, Fibrinogen |
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Improves the visualization of more protein features |
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Provides identification of many (often hundreds) or proteins |
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2D gel before Albumin depletion |
2D gel after Albumin depletion |
Electrophoresis:
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Silver stained small scale SDS-PAGE gel. This is a BSA dilution series on a Bio-Rad Ready Gel |
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Silver stained small scale 2D-PAGE gel. This is a separation of whole white fly on a Bio-Rad Criterion Gel |
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Multiplexed image of a large scale 2D-DIGE gel |
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