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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 brain tissue.
Guanylate cyclases belong to the adenylyl cyclase class-4/guanylyl cyclase family. There are two forms of guanylate cyclase, a soluble form (GCS or sGC), which act as receptors for nitric oxide and a membrane-bound receptor form (GC), which are peptide hormone receptors. The GC-C protein is composed of an extracellular domain, a single transmembrane domain, and a cytoplasmic region consisting of a kinase-like domain and a catalytic domain. It is expressed as two differentially glycosylated forms, a 130 kDa precursor form present in the endoplasmic reticulum and a 145 kDa form present on the plasma membrane. Ligand binding to the extracellular domain of GC-C promotes the accumulation of cGMP. GC-C acts as the receptor for heatstable enterotoxins, small peptides secreted by some pathogenic strains of E. coli that cause severe secretory diarrhea. GC-C also binds to guanylin and uroguanylin peptides, which modulate renal function in response to oral salt load.
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