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Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is lung carcinoma.
An increased intracellular Ca2+ concentration induces apoptotic cell death. Transiently elevated Ca2+ concentrations are required for glucocorticoid-mediated and T cell receptor-mediated pathways, leading to T cell apoptosis. ALG-2 (for apoptosis-linked gene 2) is a Ca2+ -binding protein that participates in regulatory events occuring late in the apoptotic program and where several death signals converge. ALG-2 is a protein expressed in normal brain, and to a greater extent in ischemic brain. The ALG-2 protein contains five EF-hand-like motifs and shares homology with members of the penta EF-hand family, which includes Calpain small subunits sorcin and Grancalcin.
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