The Synaptophysin (SYN) gene is located on the short arm of the X chromosome. Mutations in this gene are associated with X-linked mental retardation (XLMR). The SYN gene product is a 38 kD glycoprotein that is localized to the membrane of synaptic vesicles. Using immunohistochemistry, synaptophysin can be identified in a range of neural and neuroendocrine tissues, including neurons of the central nervous system, cells of the adrenal medulla, thyroid and pancreatic islets. As a specific marker for these tissues, it can be used to identify tumors originating from them, such as neuroblastoma, retinoblastoma, phaeochromocytoma, carcinoid, small-cell carcinoma, medulloblastoma and medullary thyroid carcinoma.
In order to improve the quality of SYN IHC-staining, Nordic immunohistochemical Quality Control (NordiQC) Institute organized an assessment of ten different concentrated Synaptophysin antibodies from different vendors (see table below). Researchers from 243 independent laboratories used various staining protocols and automation systems to assess these antibodies. The BioGenex SYN antibody clone SNP88 was the only antibody that achieved 100% sufficient staining (optimal or good) with basic protocol settings (see table). Antibodies from other vendors led to insufficient staining in 14-21% of cases. The most frequent causes of insufficient staining with these antibodies were as follows: (1) HIER in a non-alkaline buffer, (2) low concentrations of primary antibody and (3) use of less sensitive and specific detection systems. The BioGenex clone SNP88 has been used in numerous scientific publications and has been useful for detection of SYN in Acinic cell carcinoma of breast, Merkel Cell Carcinoma, Hepatoid carcinoma of the pancreas, Thyroid Paraganglioma and many other neuroendocrine tumors.
In general, optimal synaptophysin staining required HIER in alkaline buffer, careful calibration of the antibody concentration, and use of a traditional 3-step polymer based detection system such as the BioGenex SuperSensitive Polymer HRP detection systems. Colon tissue was recommended for the positive control and demonstrated strong staining of nerve cells and at least weak but distinct cytoplasmic staining in the majority of goblet cells.
The BioGenex SYP antibody (Clone SNP88) is a mouse monoclonal antibody, IgG3 Kappa isotype and is available in concentrated or ready to use (RTU) formats.
• Concentrated: 0.5ml (MU363-5UC) or 1 ml (MU363-5UC)
• RTU (Manual) 6ml (AM363-5M)
• RTU (Xmatrx Automation): 50 tests (AX363-50D) or 200 tests (AX363-YCD)
• RTU (i6000 AUtomation): 100 tests (AM363-10M)
• SYP positive control tissue slides: 5 slides for Xmatrx (FB-363M), or for Manual/i6000 (FG-363M).
BioGenex Super Sensitive Polymer HRP Detection Systems are available for Xmatrx Elite (QD550-YCDE), Xmatrx Infinity (QD550-YCXE), i6000 (QD410-YAXE) or Manual use (QD400-60KE, QD420-YIKE and QD440-XAKE) and offer excellent sensitivity and superior cell penetration ability for intense nuclear, cytoplasmic and membrane antigen staining.
Criteria for assessing SYP staining as optimal included:
1) A strong, distinct cytoplasmic staining reaction of virtually all endocrine islet cells in the pancreas.
2) A moderate to strong, distinct cytoplasmic staining reaction of neuroendocrine cells, ganglion cells and axons of the nerve plexus in the colon.
3) A moderate to strong, distinct cytoplasmic, dot-like staining reaction of the majority of cortical epithelial cells of the adrenal gland.
4) A weak to moderate staining of the majority of goblet cells in the colon mucosa.
5) An at least moderate, distinct, cytoplasmic staining reaction of the majority of neoplastic cells of the small cell lung carcinoma, and the intestinal neuroendocrine tumor.
6) No staining of neoplastic cells in the colon adenocarcinoma.
A weak cytoplasmic staining reaction of the exocrine pancreatic epithelial cells was accepted.
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