Whole Cell Activity Assays

    1. PI Turnover Assay (IP3 Production)
    2. 125I-EGF Binding
    3. RIPA lysates
    4. [3H]Thymidine Incorporation
    5. Growth in Soft Agar
    6. Incorporation of Sterols into Intact Cells

Assays of Cell Membranes

    1. EGF Receptor Autophosphorylation
    2. Peptide-based Tyrosine Kinase Assay
    3. Soluble Ligand Binding Assay

Lipid Raft Preparations

    1. Detergent-free Percoll/Opti-Prep Method
    2. Alkaline Detergent-free Method
    3. Neutral Detergent-free Method
    4. Triton X-100 Extraction Method

Protein Analysis

    1. SDS Polyacrylamide Gel Electrophoresis
    2. Precipitation Lowry Protein Assay
    3. Epitope Tags
    4. Phosphoamino Acid Analysis
    5. Western Blotting
    6. Stripping of Western Blots
    7. Immunoprecipitation

Lipid Analysis

    1. Bligh-Dyer Separation of Lipids
    2. Extraction and Separation of Inositol Phospholipids
    3. Cholesterol Assay
    4. Staining Lipids with Primulin

Purifications and Syntheses

    1. EGF
    2. EGF Receptors
    3. 125I-EGF
    4. Sterol/Cyclodextrin Complexes

Tissue Culture

    1. Media
    2. HBSS
    3. Splitting Cells
    4. Freezing Cells

    
 

 

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