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

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

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

Protein-Protein Interactions

1. Glutaraldehyde and EGS crosslinking

2. Equilibrium micro-dialysis

3. GST-pulldown

Filter Trap Assay for Aggregates

Proteomics

1. Global quantitative proteomics

Re-Folding Assays

1. Brian Freeman’s ATPase

2. b-galactosidase

3. In-vivo refolding