Core C provides basic support for laboratory procedures as required by the individual Projects 1-4. Support? includes analytical and preparative procedures as well as the development of new procedures and? optimization of existing protocols. Analytical procedures include 1) routine protein estimations using? colorimetric techniques and UV spectroscopy; gel electrophoresis for proteins and lipoproteins, 2)? Separation of proteins using 1-dimensional (1D) polyacrylamide gel electrophoresis (PAGE) including native? PAGE, SDS, urea and isoeletric focusing (IEF); and 2-dimensional (2D) (IEF in the first dimension and SDSPAGE? in the second dimension) gel electrophoresis 3) Separation of lipoproteins using agarose gels and 4)? immunological techniques including Western blotting, dot blotting, and ELISA. A variety of formats is? available including large gels, mini-gels, tube gels, and PhastSystem gels. Lipids are analyzed by enzymatic? and non-enzymatic procedures, Thin Layer Chromatography (TLC), Gas Liquid Chromatography (GLC),? High Performance Liquid Chromatography (HPLC) and Liquid Chromatography -Mass Spectrometry (LCMS).? Preparative procedures include 1) isolation of lipoproteins, chylomicrons, very low density lipoproteins? (VLDL), low density lipoproteins (LDL) and high density lipoproteins (HDL) from human plasma by? sequential ultracentrifugation 2) Purification of proteins using chromatographic techniques including affinity? and immunoaffinity Chromatography in some cases using fast protein liquid Chromatography (FPLC).? Proteins include apolipoproteins apoB48 from chylomicrons, apoB100 from LDL, apoA-l (including single? isoforms), apoA-ll, and apoCs from HDL; recombinant proteins such as subdomains of human apoA-l, apoB? and LDL receptor; human liver fatty acid binding protein (FABP) and isoforms of intestinal FABP.? Core C also oversees the general management of the Program Project laboratories, ensures an adequate? inventory of basic supplies, supervises the proper use of common equipment and training of new users, and? guarantees compliance to all fire, radioisotope and OSHA safety regulations. Graduate students, postdoctoral? fellows, and technicians are trained in analytic procedures by Core C staff to ensure uniformity and? consistency within the Program Project.
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