Biochemistry and Protein Techniques

Boston University Medical Campus, Boston MA

Investigators

Linked publications & trials

Abstract

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