Sources/Clones
Accurate (31A3, 13C4) and Biogenesis (31A3, 13C4).
Fixation/Preparation
Immunostaining in paraffin-embedded sections is enhanced by HIER in citrate buffer at pH 6.0.
Background
Protein gene product 9.5 (PGP 9.5) is a ubiquitin carboxyl-terminal hydrolase whose gene is mapped to chromosome 4p14, spans 10 kb and contains nine exons (Edwards et al, 1991). It displays 5' features, some common to many genes and others common to neurofilament neuron-specific enolase and Thy-1-antigen gene 5' regions (Wilkinson et al, 1989). PGP 9.5 is a 27 kD soluble protein which has been shown by immunostaining in all levels of the central and peripheral nervous system, many neuroendocrine cells, in part of the renal tubule, spermatogonia and non-pregnant corpus luteum (Wilson et al, 1988). Benign and neoplastic follicular center lymphoid cells also stain for the antigen (Langlois et al, 1995). The function of PGP9.5 is currently unknown. There is some evidence from studies in glioma cell lines that the protein is maximally expressed during the growth phase and that it may play a role in glial cells during brain development, in reactive gliosis or in tumorigenesis of the glial lineage (Giambanco et al, 1991). PGP 9.5 has been demonstrated in pituitary adenoma, medullary carcinoma of thyroid, pancreatic islet cell tumor, paraganglioma, neuroblastoma, carcinoid tumors from a variety of sites and Merkel cell carcinoma (Rode et al, 1985; Gosney et al, 1995).
Applications
PGP 9.5 is distinct from neuron-specific enolase (NSE) and is largely employed as a marker of nervous and neuroendocrine differentiation. However, it is of low specificity as shown in a study of bronchial carcinomas where, like NSE, PGP 9.5 actually labeled more cases of non-small cell tumors than small cell lesions. PGP 9.5 has the advantage of producing a more intense stain with less background compared to NSE but if used as a marker of neural and neuroendocrine differentiation, it must be employed in conjunction with chromogranin and synaptophysin which are more specific markers for this purpose. Other applications of PGP 9.5 include the study of unmyelinated nerve fibers in the skin and colonic mucosa, atrial myxomas and inclusion bodies in the central nervous system (Wilson et al, 1988; Gosney et al, 1995).
Comments
Before the advent of HIER, it was recommended that fresh tissues be fixed in a solution of 95% alcohol-5% acetic acid for 2-3 h to obtain optimal results. This is no longer necessary as HIER produces marked enhancement of immunoreactivity compared to other methods of antigen unmasking, with both increase in number of positive-staining cells and increased intensity of reaction within individual cells and their processes (McQuaid et al, 1995).
References
•Edwards YH, Fox MF, Povey S, Hinks LJ 1991. The gene for human neuron specific ubiquitin C-terminal hydrolase (UCHL1, PGP9.5) maps to chromosome 4p14. Annals of Human Genetics; 55: 273-278.
•Giambanco I, Bianchi R, Ceccarelli P et al 1991. "Neuron-specific" protein gene product 9.5 (PGP 9.5) is also expressed in glioma cell lines and its expression depends on is also expressed in glioma cell lines and its expression depends on the cellular growth state. FEBS Letters; 290: 131-134.
•Gosney JR, Gosney MA, Lye M, Butt SA 1995. Reliability of commercially available immunocytochemical markers for identification of neuroendocrine differentiation in bronchoscopic biopsies of bronchial carcinoma. Thorax; 50: 116-120.
•Langlois NE, King G, Herriot R, Thompson WD 1995. An evaluation of the staining of lymphomas and normal tissues by the rabbit polyclonal antibody to protein gene product 9.5 following non-enzymatic retrieval of antigen. Journal of Pathology; 175: 433-439.
•McQuaid S, McConnell R, McMahon J, Herron B 1995. Microwave antigen retrieval for immunocytochemistry on formalin-fixed, paraffin-embedded post-mortem CNS tissue. Journal of Pathology; 176:207-216.
•Rode J, Dhillon AP, Doran JF et al 1985. PGP 9.5, a new marker for human neuroendocrine tumors. Histopathology; 9: 147-158.
•Wilkinson KD, Lee KM, Deshpande S et al 1989. The neuron-specific protein PGP 9.5 is a ubiquitin carboxyl-terminal hydrolase. Science; 246: 670-673.
•Wilson PO, Barber PC, Hamid QA et al 1988. The immunolocalization of protein gene product 9.5 using rabbit polyclonal and mouse monoclonal antibodies. British Journal of Experimental Pathology; 69: 91-104.
Bibliografia
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
Accurate (31A3, 13C4) and Biogenesis (31A3, 13C4).
Fixation/Preparation
Immunostaining in paraffin-embedded sections is enhanced by HIER in citrate buffer at pH 6.0.
Background
Protein gene product 9.5 (PGP 9.5) is a ubiquitin carboxyl-terminal hydrolase whose gene is mapped to chromosome 4p14, spans 10 kb and contains nine exons (Edwards et al, 1991). It displays 5' features, some common to many genes and others common to neurofilament neuron-specific enolase and Thy-1-antigen gene 5' regions (Wilkinson et al, 1989). PGP 9.5 is a 27 kD soluble protein which has been shown by immunostaining in all levels of the central and peripheral nervous system, many neuroendocrine cells, in part of the renal tubule, spermatogonia and non-pregnant corpus luteum (Wilson et al, 1988). Benign and neoplastic follicular center lymphoid cells also stain for the antigen (Langlois et al, 1995). The function of PGP9.5 is currently unknown. There is some evidence from studies in glioma cell lines that the protein is maximally expressed during the growth phase and that it may play a role in glial cells during brain development, in reactive gliosis or in tumorigenesis of the glial lineage (Giambanco et al, 1991). PGP 9.5 has been demonstrated in pituitary adenoma, medullary carcinoma of thyroid, pancreatic islet cell tumor, paraganglioma, neuroblastoma, carcinoid tumors from a variety of sites and Merkel cell carcinoma (Rode et al, 1985; Gosney et al, 1995).
Applications
PGP 9.5 is distinct from neuron-specific enolase (NSE) and is largely employed as a marker of nervous and neuroendocrine differentiation. However, it is of low specificity as shown in a study of bronchial carcinomas where, like NSE, PGP 9.5 actually labeled more cases of non-small cell tumors than small cell lesions. PGP 9.5 has the advantage of producing a more intense stain with less background compared to NSE but if used as a marker of neural and neuroendocrine differentiation, it must be employed in conjunction with chromogranin and synaptophysin which are more specific markers for this purpose. Other applications of PGP 9.5 include the study of unmyelinated nerve fibers in the skin and colonic mucosa, atrial myxomas and inclusion bodies in the central nervous system (Wilson et al, 1988; Gosney et al, 1995).
Comments
Before the advent of HIER, it was recommended that fresh tissues be fixed in a solution of 95% alcohol-5% acetic acid for 2-3 h to obtain optimal results. This is no longer necessary as HIER produces marked enhancement of immunoreactivity compared to other methods of antigen unmasking, with both increase in number of positive-staining cells and increased intensity of reaction within individual cells and their processes (McQuaid et al, 1995).
References
•Edwards YH, Fox MF, Povey S, Hinks LJ 1991. The gene for human neuron specific ubiquitin C-terminal hydrolase (UCHL1, PGP9.5) maps to chromosome 4p14. Annals of Human Genetics; 55: 273-278.
•Giambanco I, Bianchi R, Ceccarelli P et al 1991. "Neuron-specific" protein gene product 9.5 (PGP 9.5) is also expressed in glioma cell lines and its expression depends on is also expressed in glioma cell lines and its expression depends on the cellular growth state. FEBS Letters; 290: 131-134.
•Gosney JR, Gosney MA, Lye M, Butt SA 1995. Reliability of commercially available immunocytochemical markers for identification of neuroendocrine differentiation in bronchoscopic biopsies of bronchial carcinoma. Thorax; 50: 116-120.
•Langlois NE, King G, Herriot R, Thompson WD 1995. An evaluation of the staining of lymphomas and normal tissues by the rabbit polyclonal antibody to protein gene product 9.5 following non-enzymatic retrieval of antigen. Journal of Pathology; 175: 433-439.
•McQuaid S, McConnell R, McMahon J, Herron B 1995. Microwave antigen retrieval for immunocytochemistry on formalin-fixed, paraffin-embedded post-mortem CNS tissue. Journal of Pathology; 176:207-216.
•Rode J, Dhillon AP, Doran JF et al 1985. PGP 9.5, a new marker for human neuroendocrine tumors. Histopathology; 9: 147-158.
•Wilkinson KD, Lee KM, Deshpande S et al 1989. The neuron-specific protein PGP 9.5 is a ubiquitin carboxyl-terminal hydrolase. Science; 246: 670-673.
•Wilson PO, Barber PC, Hamid QA et al 1988. The immunolocalization of protein gene product 9.5 using rabbit polyclonal and mouse monoclonal antibodies. British Journal of Experimental Pathology; 69: 91-104.
Bibliografia
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
