Chapter title |
Immunohistochemical Detection of Progranulin (PGRN/GP88/GEP) in Tumor Tissues as a Cancer Prognostic Biomarker
|
---|---|
Chapter number | 8 |
Book title |
Progranulin
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8559-3_8 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8557-9, 978-1-4939-8559-3
|
Authors |
Ginette Serrero, David Hicks, Serrero, Ginette, Hicks, David |
Abstract |
Immunohistochemistry is a well characterized and robust staining technique for the identification and localization of proteins in biological tissues. The following methodology describes the process for identifying progranulin (PGRN/GP88/GEP) in the cellular cytoplasm of target tissues such as tumor biopsies. Such tissue is collected from cancer patients and stabilized using formalin-fixed paraffin embedding after which 4-5 μm sections of the tissue are mounted on positively charged microscope slides. After deparaffinization using ethanol, the tissue is processed through an antigen retrieval process using citric acid at pH 6.4. A monoclonal antibody (Clone AG03) against human PGRN/GP88/GEP is then contacted with the tissue before a horseradish peroxidase (HRP) conjugated antibody is employed to detect AG03 bound to PGRN/GP88/GEP. Subsequent application of the HRP substrate results in color generation. Using a light microscope, a trained user can "score" the percentage of cells stained and intensity of such staining. Scoring ranges from 0 (no staining) to 3+ (strong cytoplasmic staining in >10% tumor cells). When scoring is correlated to patient outcome, we have demonstrated that a patient with tumor PGRN/GP88/GEP staining of "3+" has a fivefold increased risk of disease recurrence within 5 years. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 5 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Professor | 1 | 20% |
Librarian | 1 | 20% |
Student > Bachelor | 1 | 20% |
Student > Master | 1 | 20% |
Unknown | 1 | 20% |
Readers by discipline | Count | As % |
---|---|---|
Medicine and Dentistry | 3 | 60% |
Engineering | 1 | 20% |
Unknown | 1 | 20% |