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NORC Adipocyte Biology and Molecular Nutrition Core Services Request for Core Use Requestor Name: Campus Box: Date of Request: Phone: Fax: E-mail: Summary of Project: Funding Source (Agency/Number): PI: PLEASE ATTACH A COPY OF HUMAN AND/OR ANIMAL STUDY PROTOCOL APPROVAL LETTER(S) Questions please contact: Terri Pietka (362-8469; [email protected]) or Nada Abumrad (747-0348; [email protected]) Enter the approximate number of analyses needed for each service below. a. Cells for Culture: 3T3-L1 ____ 3T3-F442A ____ OP9 ____ HIB1B _____ LS14 _____ LiSa-2 _____ SBGS _____ C2C12 ____ HSMM ____ b. Adipose Tissue Morphology: Cell Size: ____ Cell Number: ____ c. Gene Expression Analyses: RNA Extraction? _______ Number of Samples: _________ Type of Tissue: ________ Pathways of Interest: Autophagy ECM/Fibrosis FA Metabolism Angiogensis Glucose Metabolism Inflammation ER Stress Other (Please describe below) RT-PCR details (e.g. genes to measure, tissues, etc.): Total number of genes requested: ____________ d. Protein Expression Analyses: Lysate Preparation? _______ Number of Samples: _________ Type of Tissue: ________ Pathways of Interest: mTOR Signaling ER Stress Insulin Signaling Inflammation Other (please describe below) Western Blotting details (e.g. proteins to measure, tissues, etc.: Total number of proteins to measure: _______________ e. Substrate Metabolic Assays Gucose uptake_______ Glycolysis______ Glucose Oxidation ______ Glycogenesis______ Fatty Acid Uptake______ Fatty Acid Oxidation______ Lipid Incorporation______ Lipolysis______ f. Mitochondrial Physiology: Type of analyses: isolated mitochondria ________ Tissue _________ Tissue type _________ Substrate/inhibitors requested: Glutamate/Malate______ Oligomycin ______ Pyruvate ______ Rotenone ______ Succinate _____ Antimycin A______ Uncoupling _______ Number of samples to be measured _______ g. Microscopy/Cell Imaging: Briefly describe measurements needed: h. Training: Biochemical Characterizations (circle assay(s)): Glucose uptake Glycolysis Glucose Oxidation Glycogenesis Fatty Acid Uptake Fatty Acid Oxidation Lipid Incorporation Lipolysis Triglycerides Cellular trafficking and imaging Autophagy Analysis Specifics: i. Consultation: Brief project description: Mitochondrial Function/Physiology (circle assay(s)): Respiration ROS Production Oxidative Stress ATP Production Mitochondrial Membrane Potential Mitochondrial Calcium Uptake j. Equipment Usage: ABI-7500 Fast _____ LiCor Odyssey _____ Tissue Culture Facility _____ Nikon TE2000U Microscope _____ Spectroscopy (Bio-Tek/NanoDrop) _____ Oxygraph 2K _____ Qiagen TissueLyser II ______ Special instructions, comments, etc. for any of the above services: PLEASE CITE: NORC GRANT DK056341 IN ALL PUBLICATIONS RESLULTING FROM THIS EFFORT. Return completed form to: Terri Pietka (362-8469; [email protected]) or Nada Abumrad (747-0348; [email protected]) Campus Box 8031 Approval: ______________ Date: _______________ Priority: ________________