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Welcome to the knowledge base of Ucan-U. Ucan-U wants to use this wiki to share relevant information related to its work. Currently Ucan-U only shares standard operating procedures in this system, but with your help we hope to be able to collect and organise a wealth of information in here. ... read more

Isolation of Human Mononuclear Cells from Blood

This protocol describes a method for isolating mononuclear cells in high yield and purity from small or large volumes of human peripheral blood, using a simple and rapid centrifugation procedure based on the method developed by Bøyum. Isolation of Human Mononuclear Cells from Blood ... read more

Proliferation of Human Mononuclear Cells with 3H-Thymidine

When an antigen or mitogen is presented by antigen presenting cell (APC), specific T-cells can recognize and respond to these antigens or mitogens. They will start dividing (proliferation) and can also release specific cytokines and chemokines. The proliferation can be detected by adding 3H thymidine to the dividing cells. This ... read more

Immunofluorescent staining and flow cytometric analysis of cells

Flow cytometry is a useful tool for simultaneously measuring multiple physical properties of individual particles (such as cells). Using this flow cytometric analysis protocol, one can perform a simultaneous analysis of surface and intra-cellular  molecules at the single-cell level. Immunofluorescent staining and flow cytometric analysis of cells ... read more

Thawing Human Mononuclear Cells

Viability of thawed cells depends on the quality of the cell preparation and the handling of cells prior to and during the freeze-thaw process.This protocol describes a procedure for cryopreservation of PBMC. Thawing Human Mononuclear Cells ... read more

Proliferation of Human Mononuclear Cells with CFSE

The stable incorporation of the intracellular fluorescent dye 5-(and –6)- carboxylfluorescein diacetate succinimidyl ester (CFSE) into lymphocytes is a powerfool tool to monitor lymphocyte migration in vivo and to quantitatively analyze cell divisions both in vivo and in vitro. Proliferation of Human Mononuclear Cells with CFSE ... read more

Lysis of Whole blood and Human Mononuclear Cells from Blood and Synovial Fluid

This protocol describes a lysis protocol for whole blood and human mononuclear cells from blood and synovial fluid. Lysis of Whole blood and Human Mononuclear Cells from Blood and Synovial Fluid ... read more

General Annexin V Staining Procedure

PE Annexin V is used to quantitatively determine the percentage of cells within a population that are actively undergoing apoptosis. General Annexin V Staining Procedure ... read more

Isolation of Human Mononuclear Cells from Synovial Fluid

This protocol describes a method for isolating mononuclear cells in high yield and purity from small or large volumes of human synovial, using a simple and rapid centrifugation procedure based on the method developed by Bøyum. Isolation of Human Mononuclear Cells from Synovial Fluid ... read more

Detection of proteins of interest in cell lysates

When cells are subjected to any kind of change in its direct environment (activation, inhibitor treatment, proliferation, differentiation, etc) the protein expression profile in cells changes. With Western blotting it is possible to detect the protein of interest and compare its expression profile in different circumstances. Detection of proteins of ... read more

Generation of EBV-immortalized B cell lines

Immortalization of B lymphocytes by EBV is an effective procedure for inducing long-term growth of certain human B lymphocytes. Generation of EBV-immortalized B cell lines ... read more

Immunofluorescent staining and flow cytometric analysis of human CD4+ Regulatory T cells

The following protocol describes a method to Immunofluorescent stain flow cytometric analysis of human CD4+ Regulatory T cells. Immunofluorescent staining and flow cytometric analysis of human CD4+ Regulatory T cells ... read more

Serum Isolation from Human Peripheral Blood

The following protocol describes a method for serum isolation from Human peripheral blood. Serum Isolation from Human Peripheral Blood ... read more

Plasma Isolation from Human Peripheral Blood

The following protocol describes a method for plasma isolation from Human peripheral blood. Plasma Isolation from Human Peripheral Blood ... read more

      
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