A searchable database of core laboratory facilities. Core Facilities

Molecular and Cellular Biochemistry Core (BCH)

Director: Steen, Hanno, Ph.D.; Cohen, David, M.D., Ph.D.

Core Summary:

The Molecular and Cellular Biochemistry Core is dedicated to providing HDDC investigators and their trainees with access to a variety of methods to determine the compositions of tissues, as well as their cellular and subcellular specimens. The Core Scientists offer a broad range of analyses and preparatory services including Proteomics. Importantly, the Core also provides training in the application of state-of-the-art techniques in molecular analysis of proteins, lipids and nucleic acids. The main Core laboratory is located at Brigham and Women’s Hospital (BWH) and the satellite Core laboratory is located at Children’s Hospital
Boston (CHB).

We perform a broad array of biochemical and analytical assays that are not typically available within laboratories of HDDC members. These have generally been adapted so that they may be performed in large numbers on microplates. These include quantification of lipids, mRNA and protein and a full array of mass
spectroscopy services for protein analysis (and soon to be expanded for lipidomics). We also offer both analytical and preparatory separations by a variety of methods.

Institutions:
  • Boston Children's Hospital
Personnel/Contact Information:

Director: Steen, Hanno, Ph.D.
Phone: (617) 919-2629
Email

Director: Cohen, David, M.D., Ph.D.
Phone: (617) 525-5090
Email

Facilities and Equipment:

Major Equipment:

  • FPLC apparatus
  • HPLC apparatus fitted with autosampler and UV/vis detector
  • Microplate reader
  • Nanoflow liquid chromatography/electrospray ionization mass spectrometry system
  • Quantitative real-time PCR apparatus
Services:
  • Biliary lipid concentrations analysis

    The following analyses are available:
    * Enzymatic lipid analyses of cholesterol, triglycerides, free cholesterol
    * Total Bile Acid
    * Total Phospholipids
    * HPLC analyses

  • Biliary particle analysis

    Material separated by FPLC plus cost of bile salts required for pre-equilibration of column.

    Lipid assays of fractions available as requested.

  • Fluorescence labeling of lipoproteins

    "Standard fluorescence labeling reaction for lipoproteins."

  • Lipoprotein analysis

    "Samples from plasmid of cell culture media separated by FPLC.

    The following analyses are available:
    * Lipid assays of fractions
    * Western blot analysis of fractions
    * Discontinuous gradient ultracentrifugation; can include desalting of fractions
    * Agarose gel electrophoresis of fractions"

  • Preparation of recombinant proteins

    Includes "vector design, large-scale prokaryotic or eukaryotic expression and purification." Also:

    * Production of recombinant protein by Baculovirus expression system.
    * Production of recombinant protein by E. coli.

    Price tailored to scope of project and materials.

  • Protein Identification from complex mixtures

    "In solution digestion of proteins, no further fractionation."

  • Protein identification from gel bands

    Cut bands can be:
    * Coomassie-stained
    * Silver-stained

  • Purification of biliary lipid aggregatesfrom model biles

    "Purified lipid biliary particles performed by FPLC fractionation plus cost of bile salt for column equilibration."

  • Quantitative real-time PCR

    "Tissue or cellular mRNA analysis. Nucleic acids analysis in the form of quantitative real-time PCR using a state-of-the-art, high throughput instrument. Can perform mRNA extraction plus cDNA synthesis or just real-time PCR alone depending on samples provided."

  • Radiolabeling of lipoproteins

    "Standard radiolabeling reaction for lipoproteins."

  • Sequential ultracentrifugation for lipoproteins

    Purified lipoprotein fractions with quantity scaled to investigator’s need.

  • Total lipid analysis

    Total lipid analysis can be done on:
    * Tissue or Cells
    * Plasma or cell culture media


Departmental Web Link(s):

Last updated: 2015-08-25T11:32:15.540-05:00