Protocols

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