Journal: The Journal of Experimental Medicine

Article Title: Dendritic Cells Retrovirally Transduced with a Model Antigen Gene Are Therapeutically Effective against Established Pulmonary Metastases

doi:

Figure Lengend Snippet: Allogeneic MLR of bone marrow–derived DCs and splenocytes. Bone marrow cells were transduced by coculture with retroviral producer lines, CreLacZ (DC–β-gal) and CreNeu (DC–Neu), and differentiated into DCs in vitro. DCs were cocultured with allogeneic, C57BL/6 T cells, isolated from bulk splenocytes by passing cells through an immunoaffinity column. After 3.5 d in culture, cells were pulsed with [ 3 H]thymidine as described in Materials and Methods. Results from triplicate wells were corrected for [ 3 H]thymidine incorporation by irradiated stimulators and T cells alone, and are plotted as the mean ± SEM.

Article Snippet: Bone marrow–derived DCs (transduced and nontransduced) or freshly prepared splenocytes were irradiated (2,000 rads) and plated in graded doses with 2 × 105 allogeneic T cells in 200 μl of RPMI-1640 media supplemented with 10% heat-inactivated fetal calf serum, 2 mmol/liter glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin, 1× nonessential amino acids, 1 mM sodium pyruvate (all from Biofluids), 1.25 μg/ml amphotericin B (Fungizone; GIBCO BRL ), and 50 μg/ml gentamicin sulfate ( GIBCO BRL ) (mCM) in flat-bottomed 96-well tissue culture plates and incubated at 37°C, 5% CO2 for 3.5 d. Allogeneic T cells were prepared from C57BL/6 mice by passing RBC-depleted splenocytes through an immunoaffinity column (R & D Systems, Minneapolis, MN).

Techniques: Derivative Assay, In Vitro, Isolation, Irradiation

Journal: Journal of Proteome Research

Article Title: Stable Isotope Dilution Multidimensional Liquid Chromatography-Tandem Mass Spectrometry for Pancreatic Cancer Serum Biomarker Discovery

doi: 10.1021/pr800904z

Figure Lengend Snippet: General workflow for processing of serum samples. Pooled serum from pancreatic cancer patients or controls is mixed with SILAC labeled conditioned media from CAPAN-2 cells. Abundant proteins are removed by immunoaffinity depletion. Samples were separated by IEF. IPG strips were then cut into pieces and digested with trypsin. Tryptic peptides were separated by strong cation exchange, then analyzed by reversed-phase LC−MS/MS.

Article Snippet: As expected, only a negligible amount of protein is lost from SILAP standard samples processed alone using this immunoaffinity column.

Techniques: Labeling, Electrofocusing, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry