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257 ʵÑéËÄÊ®ËÄ Á÷ʽϸ°ûÒǼì²â¾ÞÊÉϸ°û°ûÄÚϸ°ûÒò×Ó 262 µÚ°Ëƪ ÌÇÃâÒß¼¼Êõ ʵÑéËÄÊ®Îå IgGÌÇÁ´¼ì²â 271 ʵÑéËÄÊ®Áù ÃâÒßÇòµ°°×A1ÌÇÁ´µÄ¼ì²â 275 ʵÑéËÄÊ®Æß Ï¸°û±íÃæNÌǵ°°×µÄ´úл±ê¼Ç¼°¼ì²â 280 ʵÑéËÄÊ®°Ë °ûÄÚO-GlcNAcÌǵ°°×µÄ±ê¼Ç¼°¼ì²â 284 Ö÷Òª²Î¿¼ÎÄÏ× 289 ¸½Â¼ 293 ¸½Â¼Ò» ³£ÓÃÊÔ¼ÁºÍÅàÑøÒº 294 ¸½Â¼¶þ ʵÑé³£ÓÃСÊóµÄƷϵ¼°Ä£ÐÍ 307 Contents Chapter One Antigen and Antibody Reaction in vitro 1.1 Classical Antigen and Antibody Reaction in vitro 3 1.1.1 Agglutination Reaction 7 Exp.1 Direct Agglutination Reaction ¡ª Slide Agglutination Test for Blood Type Detection 7 Exp.2 Indirect Agglutination Reaction ¡ª Latex Agglutination Test for Detection of RF 12 Exp.3 Indirect Agglutination Inhibition Reactions ¡ª Indirect Latex Agglutination Inhibition Test for the HCG Pregnancy Test 16 1.1.2 Precipitation Reaction 20 Exp.4 Countercurrent Electrophoresis 20 Exp.5 Immunoelectrophoresis 25 1.1.3 Antigen and Antibody Reaction Involved by Complement 29 Exp.6 Assay of Complement Lectin Pathway 29 Exp.7 Hemolytic Plaque Assay 35 1.2 Assay of Functional Cytokine and Immunoglobin 39 Exp.8 Human Serum Specific IgE (SIgE) Assay for Anaphylaxis 42 1.3 HLA Typing Techniques 45 Exp.9 HLA Typing for B27 with PCR SSP 49 Chapter Two Isolation of Immune Cells 2.1 Separation and Purification of Immune Cells 55 Exp.10 Separation of Mononuclear Cells from Whole Peripheral Blood 58 Exp.11 Preparation of Mouse Bone Marrow Derived Macrophages 62 Exp.12 Isolation of Mouse Peritoneal Macrophages 66 Exp.13 Generation Isolation of Dendritic Cells 71 Exp.14 Isolation of Natural Killer Cells 77 Exp.15 Isolation of T Cell from Mouse Spleen Using Magnetic Beads 85 Exp.16 Isolation and Purification of B Cells by Nylon Wool 91 Exp.17 Isolation and Purification of B Cells by MACS 96 Exp.18 Isolation and Purification of Murine Alveolar Macrophages 100 Exp.19 Isolation of Immune Cells from the Mouse Intestinal Immune System 107 Chapter Three Assay for the Functions of Immune Cells Exp.20 Mouse Spleen NK Cells Activity Assay 118 Exp.21 Erythrocyte Rosette Forming Cell Test (ERFC) for Guinea Pig T Cells 123 Exp.22 Detection of T Lymphocyte Subgroups in Peripheral Blood by the SAP 127 Exp.23 One way Mixed Lymphocyte Reaction 132 Exp.24 Measurement of CTL Activity 137 Chapter Four Preparation of Antibody Exp.25 Preparation of Polyclonal Antibody 145 Exp.26 Purification of Polyclonal Antibody 150 Exp.27 Preparation of Monoclonal Antibody 157 Exp.28 Generation of scFv 167 Chapter Five Animal Models of Immuno related Diseases Exp.29 Model of Anaphylaxis Shock in Guinea Pig 178 Exp.30 Murine Model of Type ¢ñ Hypersensitivity Reaction¡ªPassive Cutaneous Anaphylaxis Model 183 Exp.31 Guinea Pig Type ¢ò Hypersensitivity Reaction¡ªForssman Cutaneous Vasculitis Response 188 Exp.32 Rabbit Type ¢ó Hypersensitivity Reaction¡ªArthus Reaction 192 Exp.33 Mouse Type ¢ô Hypersensitivity Reaction¡ªDinitrofluorobenzene induced Delayed Type Hypersensitivity 196 Exp.34 Induction of Mouse Experimental Autoimmune Encephalomyelitis 201 Chapter Six Immunolabeling Technique Exp.35 Enzyme Linked Immunosorbent Assay (ELISA) 209 Exp.36 Indirect Immunofluorescence Assay 214 Exp.37 HBsAg Test Trip by Double Antigen Sandwich Immune Colloidal Gold Strip Technique 218 Exp.38 Detection of HER 2 Extracellular Domain of Breast Cancer Patients by Chemiluminescence Immunoassay 225 Exp.39 Bioluminescence Assay 232 Chapter Seven Flow Cytometry and Cell Apoptosis Detection Technique Exp.40 Thymocytes Apoptosis Assay by Agar