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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 breast carcinoma.
GRP94 is a 803 amino acid protein belonging to the heat shock protein 90 family. It acts as a molecular chaperone that functions in the processing and transport of secreted proteins. GRP94 and its N-terminal fragment stimulates CTL expansion and maturation of human monocyte-derived dendritic cells (MDDC). It plays a central role in innate as well as acquired immunity, maturation and chemotaxis of dendritic cells, Ab production, cross-priming, as a potential marker in breast cancer and is a peptide acceptor in endoplasmic reticulum and an accessory to peptide loading of MHC class I molecules. Expression of GRP94 suppressed A23187-induced apoptosis and stabilized calcium homeostasis. GRP94 is expressed in melanoma or liver metastases of colon carcinoma cells, human gastric carcinoma BGC-823 cells.
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