Protein Biochemistry

1. Protein Expression and Purification

   1. Bacterial overexpression

   2. FPLC guidelines

   3. DEAE guidelines

   4. In-Vitro Translation

      1. Invitro Translation

      2. Nuclease Treatment of Rabbit Lysate

      3. Coupled translation/transcription

   5. Proteins

      1. Hsp70 columns

      2. Brian Freeman’s hsp70

      3. Jaewan Song’s hsp70

      4. Hip

      5. Hdj-1

      6. GST-Bag-1mm

      7. C. elegans CHIP

2. Protein Analysis

   1.  Electrophoresis

      1. SDS-PAGE (denaturing)

      2. Native SDS-PAGE/SOD active stain using native gel

      3. 2-dimensional

      4. Blatter gel

   2.  Antibodies

      1. Table of antibodies

      2. Western blot analysis using ECL

      3. Semi-Dry Transfer

      4. Western Blot Analysis Using Alkaline Phosphatase

      5. Western Blot Using Licor Odyssey Analysis

      6. Immunoprecipation

         1. general

         2. HSF1 and HSF2

      7. Immunohistochemistry

   3. Bio-rad assay for quantification

3. DNA-Protein Interactions

   1. Nuclear extracts

   2. Whole cell extracts

   3. Gel mobility shift analysis

   4. Super Shift

   5. 32P labeling of oligonucleotide probe

   6. In-vitro footprinting methods

      1. Genomic DNA isolation

      2. Ligation mediated PCR

   7. ChIP

4. Protein-Protein Interactions

   1. Glutaraldehyde and EGS crosslinking

   2. Equilibrium micro-dialysis

   3. GST-pulldown

5. Filter Trap Assay for Aggregates

6. Proteomics

   1. Global quantitative proteomics

7. Re-Folding Assays

   1. Brian Freeman’s ATPase

   2. b-galactosidase

   3. In-vivo refolding