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Biochemistry/Pharmacology Resource Laboratory

The Biochemistry/Pharmacology Resource Labs (BPRL) serve as an integrated facility where faculty can conduct investigations of mammalian cell biochemistry, physiology, and pharmacology. Individual laboratories are equipped with state-of-the-art instrumentation for rapid and concise analyses of a variety of biological materials. Current activities of the BPRL include: quantitative analyses of drugs, steroid and peptide hormones, vitamins, antibiotics, proteins, lipids, and enzymes; isolation and analyses of subcellular components (microsomes, endoplasmic reticulum, DNA, RNA); and evaluations of cell membrane and receptor function (radioligand binding, free radical biochemistry). Tissue culture facilities are available for studies requiring the growth and propagation of mammalian cells and include cell-line banking. Services offered include PCR, RT-PCR and western blots.

Laboratory specialists have expertise in biochemical pharmacological techniques in mammalian systems, provide the technical expertise necessary to operate BPRL equipment, and are available to assist investigators as necessary. We help train all graduate students in any of the above mentioned methods and on safety.

In addition to performing routine biochemical analyses for investigators throughout the college, lab specialists are involved in development and implementation of new and innovative approaches in the study of various research problems. The BPRL participates fully in the CVM Quality Assurance Program and protocols can be developed which meet Good Laboratory Practice (GLP) standards.

A. Instrumentation

  • Polytron Homogenizer - For homogenization, dispersion, fiberization, and emulsification of any type of tissue or any solid in a liquid medium.
  • High Performance Liquid Chromatography - Purification and detection of enzymes, antibiotics, proteins, vitamins, hormones, drugs, and pesticides.
  • High Speed Ultracentrifuges - Separation of various cellular components and isolation of nucleic acids, mitochondria, and membrane fractions.
  • Spectrophotometers - Fixed wavelength or scanning spectral analysis, quantitation, kinetic studies.
  • Gel Electrophoresis Equipment - Equipment to run PAGE gels and two-dimensional gels.
  • Vacuum Pumps - For filtration and drawing fluids through columns.
  • Microscopes, Phase Contrast, and Fluorescence, Upright and Inverted Characterization of cellular activity.
  • Scintillation, Gamma Counters - Quantitation of isotope uptake and radioimmunoassay data.
  • Tissue Culture Instrumentation - Laminar flow hoods, CO2 incubators for sterile tissue culture and dispersed cell culture work.
  • Freezers (—70°C) - Help stop tissue degradation.
  • Cell Disrupter, Ultrasonic, ‘Sonicator’ - Designed for laboratory cell disruption; can prepare an emulsion down to 0.01 microns, homogenize 'immiscible' liquids, polymerize some materials, depolymerize others, and accelerate chemical reactions.
  • Flexi-dry Lyophilizer - Used for most standard freeze-drying procedures.
  • Convection Oven - Controllable operating temperate range is 75°C to 225°C.
  • UV Max Microplate Reader - Reads all 96 wells in 5 seconds; good for protein assays, enzyme kinetics, analyses.
  • Fluorescence Microplate Reader - Quantitates uptake of fluorescent dyes in or on live cells; biochemical assays.
  • Shaking Water Bath - Precise temperature control within the temperature range of -10°C to 100°C; oscillating speed is 20 to 200 oscillations per minute.
  • Ultrasonic Cleaner - Fast and safe cleaning method for almost any type of small items.
  • Sample Concentrator and Dry Block Units - Designed to accelerate the concentration by evaporation of sample solutions prior to analysis.
  • Microphysiometer - Measures extracellular acidification rates in response to externally applied reagents.

B. Technical Expertise

  • Kinetics and purification and characterization of enzymes (total glutathione, super oxide dismutase, and neurotoxic esterase).
  • Tissue and cell culture (bovine pulmonary endothelial cells, viability studies, response to toxicants).
  • Radio-immunoassays for hormones (LH, FSH, T4, ACTH, cyclic AMP and prostaglandins, insulin, sex hormones).
  • Radiotracer studies (tritiated leucine in mouse embryos and cytotoxicity tests).
  • ELISA assay of GFAP and S-100B
  • Analytical chemistry and detection of metabolites (dye binding protein assay, carbaryl and aldicarb metabolites).
  • HPLC of pesticides or antibiotics in tissues.
  • Electrophoresis and separation of macromolecules (proteins and nucleic acids).
  • Quantitative analysis of drug or xenobiotics in biological materials (heavy metals and pesticides).
  • Chromatography (HPLC of polyamines, insecticides residues, drugs [rifampin], and oxytocin).
  • Spectrophotometry (detection of proteins and other quantities in biological samples).
  • Toxicology (NTE, Cholinesterase, protein assays on organophosphates, functional observation battery [FOBI on rat behavior).
  • Injection of rabbits and mice IV for antibody production, antibody purification; Blood collecting from rats, mice, and poultry.
  • Preparation of buffers, solutions, gels, and culture media.
  • Development of primary cell cultures (equine endothelial cells).
  • Determining covalent binding of OPs to muscarmnic receptor sites in human neuronal cells.
  • PCR and Other Molecular Techniques, RT-PCR, northern & western blots.

C. Using the Biochemistry/Pharmacology Resource Laboratories

To use the Biochemistry/Pharmacology Resource Laboratory, please contact the Laboratory Supervisor, Delbert Jones () at 231-3718 to schedule a meeting to discuss your project needs, timelines, and associated costs.

The Faculty Laboratory Coordinator is Dr. William Huckle (231-3620).

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