Practical Immuno 2 PDF

Practical II Cellular diagnostics how to detect one cell = immunology  mb proteins CD (cluster of differentiation) T-helper cells...

Practical II Cellular diagnostics how to detect one cell = immunology  mb proteins CD (cluster of differentiation) T-helper cells model profile on the mb of the cell that is spe for this cell found which molecule is specific for this cells - CD 4 in T helper cells polyclonal = all pool of Ab is activated production of Ab against one of this protein = identification of this spe cell Production of polyclonal antibodies  Production of monoclonal antibodies identification only of one part of the molecule = harder you have to find 1. entire molecules 20 epitopes injection to other species 2. fuse this cell with the cell that is immortal = cancer cells hybridoma = all cells from the spleen and myoloma (cancer cell) only in high productive rate 1 cell of hybridoma in each tube 100 of wells of hybridoma CD (clusters of differentiation) constant region = fluorescent dye Enzymatic reaction  Others indirect Dye S+P=Signal Luminiscence fluorescence fluorescence radioactivity Flow cytometry  Flow cytometry A laser- or impedance-based, biophysical technology employed in cell counting, cell sorting, biomarker detection and protein engineering, by suspending cells in a stream of fluid and passing them through an electronic detection apparatus.  Separation Form Fluorescence colour Charge Immunohistology Extraction of Lymphocytes Lymphocytes and monocytes  Procedure FITC Anti-human CD4 Direct test to detect Helper T-cells The slides are total lymphocytes Prepare the slides 20microL of product 15 min 2X10 min PBS Mount the slides with covers Microscope Results