Chemistry & Biochemistry Core
Core Leader: Haya Herscovitz, Ph.D. (faculty page)
Core Director: Cheryl Oliva England, B.S.
Laboratory Manager: Michael S. Gigliotti, B.S.
The Chemistry & Biochemistry Core Facility provides technical support for analytical and preparative laboratory procedures, modifies/adapts/ optimizes existing protocols and develops new methodologies as required by the investigators. This support includes protein, lipid and lipoprotein purification and analysis.
The core offers the following procedures:
Separation of proteins by 1-dimensional (1D) polyacrylamide gel electrophoresis (PAGE) including native PAGE, SDS, urea and isoelectric focusing (IEF) and 2- dimensional (2D) (IEF in the first dimension and SDS-PAGE in the second dimension) gel electrophoresis. Proteins separated by 2D gel electrophoresis can then be excised from the gels and analyzed by Mass Spectrometry.
A variety of formats is available including large gels, minigels tube gels and PhastSystem gels.
Separation of lipoproteins by agarose gel electrophoresis
Analysis of proteins by immunological techniques including Western blotting and dot blotting.
Purification of proteins by liquid chromatography including High Performance Liquid Chromatography (HPLC), Fast Protein Liquid Chromatography (FPLC) and Affinity Chromatography, Heparin and immunoaffinity chromatography.
Purification of lipoproteins from human plasma by sequential and density gradient ultracentrifugation.
Purification of human apolipoproteins from HDL including single isoforms (apoA-I, apoA-II and apoCs).
Separation and identification of lipids by Thin Layer Chromatography (TLC), Gas Liquid Chromatography (GLC) and HPLC.
Purity check of lipids using TLC, GLC, HPLC and FPLC.
Quantification of lipids, triacylglycerols, free and esterified cholesterol and phospholipids by enzymatic and chemical techniques.
Detailed characterization of lipids is carried in collaboration with the Mass Spectrometry facility (Dr. C. Costello)