-
[Immunology] Antibody Purification 日期：2005-07-18 00:00:00 点击：204 好评：0
Monday, May 03, 2004 Description This protocol includes an ammonium sulfate cut, affigel blue chromatography and affinity chromatography Procedure Pour a 5 ml affigel blue column (biorad) and wash with 50 ml Prewash to prep the column (whe...
-
[Immunology] Isolation of cell nuclei for the application in the cell-fre 日期：2005-07-18 00:00:00 点击：254 好评：0
Monday, November 24, 2003 Description Pretreatment of epithelial cells with 20 M Cytochalasin B for 30 min in RPMI prior to harvesting results in the reduction of cytoskeletal microfilaments and the easier release of the nuclei from the su...
-
[Immunology] Preparation of cytoplasmic extracts for the application in a 日期：2005-07-18 00:00:00 点击：167 好评：0
Monday, November 24, 2003 Description Cells are grown to 80% confluency, then harvested, washed and disrupted in KPM buffer by freezing-thawing cycles with liquid nitrogen essentially as described by Fearnhead et al., 1997, Genes Dev. 11:...
-
[Immunology] ApoptosisReconstitution of activated cytoplasmic extracts wi 日期：2005-07-18 00:00:00 点击：210 好评：0
Monday, November 24, 2003 Description Reconstitution of activated cytoplasmic extracts with isolated radioactive nuclei. Analysis of apoptotic activity by quantitative, radioactive DNA fragmentation assays or DAPI staining. Procedure Radio...
-
[Immunology] Analysis of apoptotic activity by qualitative DNA laddering 日期：2005-07-18 00:00:00 点击：238 好评：0
Monday, November 24, 2003 Description Reconstitution of activated cytoplasmic extracts with isolated nuclei. Analysis of apoptotic activity by qualitative DNA laddering assay and DAPI staining. Procedure Reaction mix Cytoplasmic extracts w...
-
[Immunology] Activation of apoptotic activity in cytoplasmic extracts for 日期：2005-07-18 00:00:00 点击：199 好评：0
Monday, November 24, 2003 Description Activation of apoptotic activity in cytoplasmic extracts for western blot analysis Procedure Activation reaction Cytoplasmic extracts were activated according to following protocol (as an example, cyto...
-
[Immunology] Activation of apoptotic activity in cytoplasmic extracts and 日期：2005-07-18 00:00:00 点击：186 好评：0
Monday, November 24, 2003 Description Activation of apoptotic activity in cytoplasmic extracts and measurement of the resulting caspase enzymatic acticity Procedure Activation reaction Cytoplasmic extracts were activated according to follo...
-
[Immunology] ELISPOT for IFN-gamma and IL-10 日期：2005-07-18 00:00:00 点击：205 好评：0
Wednesday, October 29, 2003 Description An ELIspot assay to count cytokine-producting cells Procedure -Coat plates with primary Antibody 2 g/ml, 50 l/well -Incubate overnight 4C -Aspirate (do not remove the cover to keep the plate sterile)...
-
[Immunology] ELISA for IFN-gamma and IL-10 日期：2005-07-18 00:00:00 点击：210 好评：0
Wednesday, October 29, 2003 Description A simple ELISA assay to measure cytokines in culture supernatants Procedure -Coat plates with primary Antibody 2 g/ml, 50 l/well -Incubate overnight 4C -Wash 3 times -Block with 10% FCS, 200 ml/well...
-
[Immunology] Antigen-specific intracellular Calcium flux measurement 日期：2005-07-18 00:00:00 点击：198 好评：0
Wednesday, October 29, 2003 Description A method to measure Ag-specific Calicium flux in mice cells. After immunization of mice lymphocytes are collected and restimulated in vitro. In parallel, antigen-presenting cells are loaded with the...