An intracellular cytokine (INF-gamma) staining assay is used to analyze the function of lymphocytes at the single cell level. By combining surface staining and intracellular cytokine staining, this assay can reveal the percentage of cytokine-releasing cells in a particular population, which cannot be obtained from an ELISpot assay.

BioMag Goat Anti-Rat IgG is a standard BioMag particle coated with polyclonal goat anti-rat IgG antibodies and is highly suited for use in cell sorting methods where a rat IgG antibody is used as a primary antibody. BioMag Goat Anti-Rat IgG can be used to separate the cells of interest from a heterogeneous cell population using negative selection. BioMag Goat Anti-Rat IgG can also be used as a secondary antibody in enzyme immunoassays and radioassays that utilize a rat IgG primary monoclonal antibody.The efficiency of enrichment by negative selection depends on antigen availability and the total cell population. The author has intensive experience about enrichment of mouse CD4CD8 double negative (DN) thymocytes from total thymocytes which contains only about 2% DN population. Usually the DN population occupies more than 80% of total thymocyte after one round of enrichment. However the depletion efficiency is much lower for bone marrow cells.

Total RNA is extracted from fixed biological specimens by this method with higher yield than commercial kits. The product contains intact micro RNAs and small RNAs, and fragmented long RNAs.

Mixed neuron-glia cultures provide a unique tool to study cellular contribution and molecular pathways in various neurological disorders. They are also invaluable for exploring neuron-glia interaction under physiological and pathological conditions. The relatively long-lasting midbrain neuron-glia mixed cultures generated following this protocol have been widely used to study the pathogenesis of Parkinson’s disease, the most common neurodegenerative movement disorder.

Embryonic stem cells are derived from inner cell mass of an embryo that can differentiate into every cell type in the body. Clinically, cultured red blood cell supply is of great interest. However, some of the hurdles need to be overcome. This protocol describes a two step protocol to form embryoid body and differentiate them in to hematopoietic lineage cells, especially erythroid cells.