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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 stomach adenocarcinoma or breast carcinoma.
NGAL (Neutrophil gelatinase-associated lipocalin, lipocalin 2, siderocalin and neutrophil lipocalin) is a 25 kDa glycosylated single chain monomer and member of the lipocalin family of proteins that bind and transport small lipophilic molecules. NGAL is released by activated neutrophils, can form dimers and small amounts of higher oligomers, and complexes with matrix metalloproteinase 9 (MMP-9, gelatinase B). Low level expression of NGAL in a variety of epithelia may be increased in inflammation or cancers. Expression of NGAL in the kidney is dramatically increased by ischemic or nephrotoxic kidney injury.
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