Protein Expression

• Establishment of Stable Transfectant of CHO Lec Cells (J. Takagi)
• Transient Transfection into 293T Cells (J. Takagi)
• Preparing a Selenomethionyl Protein (S. Lindgren)

Protein Purification, Characterization

• Sepharose Affinity Gels (T. Springer)
• Purification of ICAM-1 (M. Ferzly)
• SDS Slab Gel Electrophoresis (T. Springer)
• Preparation of Tonsil Lysate (M. Ferzly)
• [Sandwich] ELISA for Detecting Secreted ICAM-1 (C-D. Jun)
• Purification of Rat Monoclonal Antibodies (T. Springer)

Radioactive Experiments

• Bench Top Radioactive Work Protocol (M. Ferzly)
• Radioiodination (J. Takagi)
• Metabolic Labeling and Immunoprecipitation (J. Takagi)
• Internal radiolabeling of monoclonal immunoglobulins (T. Springer)
• High Specific Activity 3H-labeled Ig for Cross-Inhibition experiments (T. Springer)
• Protein Iodination (T. Springer)
• Anti-Ig Indirect Assay for mAb binding to Cell Surface Antigens (T. Springer)
• Immunoprecipitation for Analysis of Radiolabeled Cell Surface Molecules (T. Springer)

• Polyacrylamide Fluorography (T. Springer)

Immunochemical Methods

• Triple Sandwich Immunoprecipitation System (F. Sanchez-Madrid)
• Immunoblotting on Nitrocellulose (F. Sanchez Madrid)
• Conjugation of protein to FITC (T. Springer)
• Semi-Quantitative Measurement of Proteins by Dot Blotting (J. Takagi)
• Purification of mAb (IgG) (C-D. Jun)
• Direct Immunofluorescence Staining of Cell Surfaces in Whole Blood (Becton Dickinson)

• Quantitative Single Radial Immunodiffusion (T. Springer)

Cell Biology Protocols

• Purification of Mononuclear Cells and Neutrophils on Density Gradients (T. Springer)
• Purification of Human Mononuclear Cells and Neutrophils (C-D. Jun)
• Separation of Platelets from Whole Blood (A. Salas)
• L-selectin-PNAd interactions under flow conditions. Flow Chamber Assay (A. Salas)
• FACS Analysis Protocols (T. Springer)
• Paraformaldehyde Fixation of Fluorescent Stained Hematopoietic Cells (Becton Dickinson)

Hybridomas, Cell Culture

• Cell Hybridization Using PEG (T. Springer)
• Freezing Cells, Liquid N2 storage (T. Springer)
• Chemotaxis Assay (M. Swope)
• Soft agar cloning (T. Springer)
• Large scale media prep (T. Springer)

Cloning/Molecular Techniques

• Lentiviral transduction (T. Schurpf)