Chromatin immunoprecipitation protocol to analyze histone modifications in Arabidopsis thaliana. Werner Aufsatz, Matzke Lab. Gregor Mendel Institute of Molecular Plant Biology, Austria. Includes Chromatin Crosslinking, Chromatin preparation, Pre-clearing and immuno precipitation (IP), Collection,washes and elution of immune complexes, Reverse crosslinking and DNA cleanup - [Read Chromatin immunoprecipitation protocol to analyze histone modifications in Arabidopsis thaliana]
Gliotoxin is a metabolite of Aspergillus fumigatus that exhibits immunosuppressive activity against certain cells of the immune system. Secretion of gliotoxin during infection has been suggested as being a factor in the pathogenesis of aspergillosis. Gliotoxin secretion can be assayed in a number of ways by thin layer chromatography (TLC) high performance liquid chromatography (HPLC) or bioassay using the effect of gliotoxin on human cells1. - [Read Method for Assaying Gliotoxin Production in Aspergillus fumigatus Protocol]
Immunoprecopitation method, the protein from the cell or tissue homogenate is precipitated in an appropriate lysis buffer by means of an immune complex which includes the antigen (protein), primary antibody and Protein A-, G-, or L-agarose conjugate or a secondary antibody-agarose conjugate - [Read Protocol Immunoprecipitation]
Transfection of primary leukocytes has traditionally been a challenging but much desired protocol. It allows not only the analysis of cells in a more natural state to a cell line system, it enables the direct comparison of, for e.g. transcriptional activity using luciferase reporters, in immune cells taken from genetically-altered mice. In addition, importantly it allows for "rescue experiments" in knockout cells & the ability to over-express or reconstitute wild-type and/or mutated constructs. - [Read Transfection of Bone Marrow-Derived Mast Cells for Transcription Factor Luciferase Reporter Assays]
Troubleshoot Immunoprecipitation. Chemicon. The most common challenge with immunoprecipitation is trying to lower the number and type of background proteins that contaminate the washed immune complexes. Suggestions for decreasing background in IP. - [Read Troubleshoot Immunoprecipitation]