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Assessing Licensing of NK Cells

Natural killer (NK) cells express receptors to detect and kill target cells based on expression of target cell surface molecules. Through a process termed NK cell licensing, only NK cells that express inhibitory receptors (e.g., Ly49 receptors in the mouse) for self-major histocompatibil ...

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Subset Analysis of Human and Mouse Mature NK Cells

Identification of natural killer (NK) cell subsets has gained attention with the recent discovery that mature mouse NK cells comprise two distinct stages. Delineation of the stages is performed using the markers CD27 and CD11b on gated NK cells. The significance of this finding is underpinn ...

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Isolation of NK Cells and NK-Like Cells from the Intestinal Lamina Propria

Being exposed to food products, pathogens and harmless commensal bacteria, the mucosal immune system faces a constant challenge. Therefore, maintenance of a homeostatic balance is required to achieve tolerance to harmless bacteria and their products and to induce potent immunity to i ...

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Analysis of Uterine Natural Killer Cells in Mice

The term uterine natural killer (uNK) cell is applied in mice to an abundant but transient NK cell population that undergoes unique, terminal differentiation within embryo implantation sites during endometrial decidualization and pregnancy. In mice, decidualization is induced by ...

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Natural Killer Cells in Human Pregnancy

Natural killer (NK) cells account for 70% of the leukocytes in the mucosal lining of the uterus (the decidua) in the first trimester of pregnancy. They are CD56superbright granulated cells expressing a repertoire of Killer-cell Immunoglobulin-like Receptors (KIR) skewed towards reco ...

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Cell-Free Bead-Based Detection of Total and Phosphorylated Proteins in Plasma and Cell Lysates: Detection of FLT3

Frequently direct measurement of proteins or their phosphorylation in intact cells is not possible, for instance, when cells are too few, frozen, or subject to degradation. We have demonstrated that tumor cells pour their DNA, RNA, and protein content into circulation because of turnover and ...

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Measuring Humanized Antibodies in Plasma of Patients Treated With Antibody-Based Therapy Using Bead-Based Flow Cytometry: The Story of Alemtuzumab

Alemtuzumab (Campath), the humanized rat monoclonal antibody that targets the CD52 surface antigen, is currently used for treatment of patients with resistant chronic lymphocytic leukemia. Monitoring levels of the antibody in plasma/serum could provide insight into the optimal ...

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Detection of Chromosome Translocations by Bead-Based Flow Cytometry

Chromosome translocations resulting in fusion genes have been implicated in leukemogenesis. The paradigm involves the fusion of the genes encoding BCR and ABL, leading to a constitutively active tyrosine kinase. The detection of BCR-ABL has been limited to fluorescence in situ hybrid ...

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Detection of the MDR1 P-Glycoprotein Expression and Function

Acquired and intrinsic multidrug resistance is the major reason for the failure of anticancer chemotherapy. The most important component of clinical multidrug resistance is mediated by P-glycoprotein (Pgp), an ABC transporter encoded by the MDR1 gene and expressed on the membrane of tu ...

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Detection of Human Antibodies Generated Against Therapeutic Antibodies Used in Tumor Therapy

Application of monoclonal antibodies (MAb) for therapeutic purpose may induce the formation of human antibodies directed against the immunogenic epitopes, which are presented on the therapeutic MAb. Formation of such human antibodies mostly is an undesired side effect, but in the case ...

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Measuring Allergen-Specific IgE: Where Have We Been and Where Are We Going

About 40 yr ago, two groups of investigators identified a new class of immunoglobulins, IgE. By exchanging their results and reagents, they proved that the immunoglobulin responsible for immediate hypersensitivity was IgE. From that day forward the science of allergy was greatly advanc ...

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Development and Characterization of Mouse Hybridomas

Cell fusion protocols that were developed by Kohler and Milstein in the mid-1970s and aimed at producing and characterization of mouse monoclonal antibodies (MAbs) remain the gold standard of hybridoma development. Despite tremendous progress in using MAbs in multiple research, dia ...

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Recombinant Monoclonal Antibodies

Recombinant antibody technology is a rapidly evolving field that enables the study and improvement of antibody properties by means of genetic engineering. Moreover, the functional expression of antibody fragments in Escherichia coli has formed the basis for antibody library gene ...

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Soluble Fc Fusion Proteins for Biomedical Research

As a source of recombinant antigen, soluble constant fragment (Fc) fusion proteins have become valuable reagents for immunotherapy and laboratory investigations. Additional applications for these reagents include flow cytometry, immunohistochemistry, and in vitro acti ...

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The Use of the Antibodies in the Diagnosis of Leukemia and Lymphoma by Flow Cytometry

Flow cytometry is an automated analysis of cells passing in the fluid suspension through a laser light beam, which react with monoclonal antibodies specific for a variety of cell surface antigens. A specimen of peripheral blood, bone marrow, or other cell suspension is incubated with fluores ...

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Quantification of Surface Antigens and Quantitative Flow Cytometry

Measuring expression levels of cell surface antigens is important for the diagnosis of diseases such as B-cell chronic lymphocytic leukemia and the monitoring of targeted therapy, particularly antibody-based therapy. In some cases, the number of antigens that the therapeutic anti ...

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CD13 and CD10 Expression of Granulocytes as Markers for the Functioning of the Immune System: Quantification of the Expression of Membrane Molecules U

HLA-DR expression on monocytes as a marker for the functioning of the immune system is known to be severely depressed in immunodeficiency. Up to now, other markers for the function of the immune system are scarce. In the peripheral blood of patients with open heart surgery the expression of the membra ...

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Monitoring Cell Signaling Pathways by Quantitative Flow Cytometry

As the signaling pathways involved in leukemogenesis are being elucidated, several proteins have emerged as potential targets for therapy. Downstream from those targets are numerous intracellular factors that are constantly modulated. Monitoring those factors could provi ...

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Antibodies and Immunohistochemical Evaluation for the Diagnosis of Hematological Malignancies: An Overview

The immunohistochemical evaluation of hematopoietic tissues has expanded our knowledge of both the function and pathological processes in a wide range of lymph nodal, extranodal tissues, and bone marrow. It is impossible to cover in detail the full range of immunohistochemical marke ...

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Bead-Based Multianalyte Flow Immunoassays: The Cytometric Bead Array System

Analytical cytometry has significant potential beyond cellular analysis. The inherent capability of flow cytometers to efficiently discriminate between uniformly sized particles based on their intrinsic properties provides the foundation for multiplex bead assays. The ...

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