T Cell Protocols: Development and ActivationKelly P. Kearse The purpose of T Cell Protocols: Development and Activation is to c- lect a series of protocols, particularly those that have been developed within the past few years, to help investigators master new techniques (or improve existing ones) for the study of T-cell Biology. Invariably, in putting together a book like this it is difficult to decide which methods to include and which to leave out. To this end methods were selected from a variety of disciplines, including cellular immunology, b- chemistry, and molecular biology, to try to provide something of interest for everyone who works on T-cell development and activation. I would like to mention that my primary reason for agreeing to put this book together is that, when I was a graduate student, I purchased a copy of Selected Methods in Cellular Immunology by Mishell and Shigii which proved a tremendous help in learning the basics of one-and two- dimensional gel te- niques (and other methods). The cover has long since fallen off, but it still remains one of my most valued reference books for the laboratory. It is my hope that T Cell Protocols: Development and Activation will prove similarly useful to current and future scientists wishing to learn new methods for expl- ing the development and activation of T cells. |
From inside the book
Results 1-5 of 39
Page vii
... Immature CD4+CD8+ Thymocytes by Panning with Anti-CD8 Antibody Terrence G. Gardner and Kelly P. Kearse 5 Transfection and Transcription of Genes in Developing Thymocytes ....................................... Alison M. Michie and Juan ...
... Immature CD4+CD8+ Thymocytes by Panning with Anti-CD8 Antibody Terrence G. Gardner and Kelly P. Kearse 5 Transfection and Transcription of Genes in Developing Thymocytes ....................................... Alison M. Michie and Juan ...
Page viii
... Immature Murine T-Lymphoma Cell Lines Ellen R. Richie and Dawn A. Walker .............................................. 15 Production and Characterization of T-Cell Hybridomas PART III . OVERVIEW FOR CHAPTERS 17–29 177 Janice White ...
... Immature Murine T-Lymphoma Cell Lines Ellen R. Richie and Dawn A. Walker .............................................. 15 Production and Characterization of T-Cell Hybridomas PART III . OVERVIEW FOR CHAPTERS 17–29 177 Janice White ...
Page 3
... immature thymocytes through several unique stages characterized by the expression of distinct combinations of cell surface markers. The advancement of thymocytes from one stage to the next requires the successful completion of one or ...
... immature thymocytes through several unique stages characterized by the expression of distinct combinations of cell surface markers. The advancement of thymocytes from one stage to the next requires the successful completion of one or ...
Page 7
... immature thymocytes through a major check- point in the early differentiation sequence (the β checkpoint). Two T-cell lineages have been identified, αβ and γδ, characterized by the type of T-cell receptor (TCR) being expressed. The αβ ...
... immature thymocytes through a major check- point in the early differentiation sequence (the β checkpoint). Two T-cell lineages have been identified, αβ and γδ, characterized by the type of T-cell receptor (TCR) being expressed. The αβ ...
Page 10
... immature B cells. T-cell development in Tcf-1-deficient thymi is blocked at the immature CD8 SP stage (36), at which DN precursors have initiated CD8 expression prior to becoming DP cells. These precursors do not proliferate and fail to ...
... immature B cells. T-cell development in Tcf-1-deficient thymi is blocked at the immature CD8 SP stage (36), at which DN precursors have initiated CD8 expression prior to becoming DP cells. These precursors do not proliferate and fail to ...
Contents
Transfection and Transcription of Genes | 55 |
and Reexpression Assay | 63 |
Recombinant Adenoviruses | 69 |
Quantitative Analysis of the Usage of Human TCell | 81 |
Flow Cytometric Detection of Cytoplasmic and Surface CD3ε | 103 |
Assessing Apoptosis of Developing T Cells by Flow Cytometry | 117 |
Isolation of TCell Subsets by Magnetic Cell Sorting MACS | 133 |
A Practical Guide | 153 |
Measurement of Lymphoproliferation at the SingleCell Level | 243 |
Digital Image Analysis of Lymphocyte Activation | 257 |
Induction of TCell Signaling by Immobilized Integrin Ligands | 265 |
Cytotoxic TCell Adherence Assay CAA | 277 |
Detecting Ubiquitinated TCell Antigen | 283 |
Measurement of Protein Tyrosine | 301 |
Protein | 307 |
Activation of Heterotrimeric GTPBinding Proteins Upon | 319 |
Production and Characterization of Immature Murine | 177 |
Production and Characterization of TCell Hybridomas | 185 |
Something Happens A Brief History of | 197 |
Measurement of Phosphatidylinositol PI Hydrolysis | 211 |
Fluorescence Polarization as an Early Measure | 221 |
Generation of Human T Helper 1 and T Helper 2 Subsets | 325 |
Transcriptional Expression Analysis of TCell Activation | 337 |
Index | 355 |
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Common terms and phrases
analysis Annexin anti-CD3 antibody antigen antigen receptor apoptosis assay beads Biology CD8+ cDNA cell lines Cell Protocols cell receptor cell surface cell suspension centrifuge clones column complex conjugated containing cytokines cytometer cytoplasmic dendritic cells detection Development and Activation differentiation dilution EDTA electroporation EtBr expression fetal filter FITC flow cytometry fluorescence gene Gibco-BRL human hybridization hybridomas immature immune Immunol Immunology immunoprecipitated Incubate integrin intracellular isolated K. P. Kearse labeled ligands lineage lymphocytes lymphoma lysis buffer measurements medium membrane MHC class mice molecules mouse murine negative selection Note PBMC pellet peptide phenotype phosphatase phosphorylation pipet plate population precursor probe procedure protein reagents Resuspend room temperature sample signaling solution specific staining sterile stimulation Subheading subsets supernatant T-cell T-cell development thymic thymocytes thymus thymus lobes tissue culture transfection transgenic Tris-HCl tube tyrosine kinases ubiquitination vector washing buffer µg/mL