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Protocols

Peter Agre and students

  

A.    Tissue Culture Protocols

 
B.    
Biochemical Assay Protocols

 
C.   
Immuno Protocols

 
D.    
Molecular Protocols

  


A.   Tissue Culture Protocols:

  1. Dissociated Primary Cortical Neuron Culture
    1. Embryonic day 18 rats
      1.  Dissociated via Trypsin
    2. Embryonic day 18 or Post-natal day 1-2 mice
      1. Dissociated via Papain
    3.  Embryonic day 18 single mouse
      1. Dissociated via Trypsin
  2.  Dissociated Primary Hypothalamic Neuron Culture
    1. Embryonic day 16-17 rats
      1. Dissociated via Papain
  3. Dissociated Primary Hippocampal Neuron Culture
    1. Embryonic day 18 rats
  4. Primary Astrocyte Cell Culture
    1. Postnatal day 1-3 rats
  5. Dissociated Primary Olfactory Receptor Neuron Culture
    1. Postnatal day 1-2 rats
    2. Postnatal day 1-2 mouse
    3. Adult human tissue
  6. Dissociated Primary Olfactory Bulb Neuron Culture
    1. Embryonic day 18 rat & mice, Postnatal day 1-2 rat & mice
  7. Olfactory Bulb and/or Olfactory Epithelial Explant Cultures
    1. Embryonic day 16 mice
  8. Cell Lines
    1. 3T3-L1A
    2. PC-12


B.  Biochemical Assay Protocols:

  1. Fatty Acid Oxidation of 14C-Palmitate
  2. Acetate Incorporation Assay - Fatty Acid Synthase Activity Assay
  3. Measurement of Intracellular ATP
  4. AMPK/pAMPK Western Blot Analysis
  5. AMPK Activity Assay Protocol - SAMS Peptide Assay
  6. Cellular Viability Assays
    1. Calcein and Propidium Iodide Assay
    2. XTT
  7. Adenylyl Cyclase Assay (with Alpha Toxin)
  8. Guanylyl Cyclase Assay

 
C.  Immunostaining Protocols:

  1. General Immunohistochemistry
  2. GFAP, NST Immunohistochemistry
  3. BRDU, NST Immunocytochemistry
  4. DAPI Staining
  5. Tunel Staining
  6. Whole Mount Staining Protocols
    1. Fluorescent Staining
    2. X-GAL Staining for Whole Mounts
    3. Other Whole Mount X-Gal Protocols
  7. OE1, GAP43 Fab Immunohistochemistry
  8. Lectin Staining
  9. NT-3 with amplification


D.  Molecular Biology Protocols:

  1. Real-Time (RT) PCR
  2. Chromatin Immunoprecipitation (ChIP)
  3. In Situ Hybridization
 
 
 
 
 
 

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