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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 10 min at room temperature. A suggested positive control is bone marrow or lymphoma.
Spleen tyrosine kinase (SYK), along with ZAP70, are cytoplasmic tyrosine kinases that are viewed as potential druge targets for allergic disorders and autoimmune diseases. SYK may also play a role in cancer. Syk translocates to the plasma membrane upon B cell antigen receptor (BCR) or the high-affinity IgE receptor (FcepsilonRI) triggering, and phosphorylates downstream adaptor proteins, thereby providing docking sites for initiation of subsequent signaling pathways, such as calcium mobilization, cytoskeleton remodeling, or transcription of specific genes. Syk binds to the receptor assemblies through interactions of its pair of SH2 domains with ITAM motives of the receptor, which have been phosphorylated by Src-family kinases. These kinases also help to activate Syk by phosphorylation of its activation loop.
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