Por favor, use este identificador para citar o enlazar este ítem: https://doi.org/10.14670/HH-30.331

Título: An effective and practical immunohistochemical protocol for bone specimens characterized by hyaluronidase and pepsin predigestion combined with alkaline phosphatase-mediated chromogenic detection
Fecha de publicación: 2015
Editorial: F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología
Cita bibliográfica: Histology and Histopathology, Vol. 30, n.º 3 (2015)
ISSN: 1699-5848
Materias relacionadas: CDU::5 - Ciencias puras y naturales::57 - Biología::576 - Biología celular y subcelular. Citología
Palabras clave: Immunohistochemistry
Bone tissue
Antigen retrieval
Resumen: The aim of this study was to provide an effective procedure for immunohistochemistry (IHC) investigations of bone specimens. Samples from rat femoral and human vertebral bone were processed with a detailed and effective IHC protocol summarized here. First, a novel antigen retrieval (AR) method of hyaluronidase combined pepsin predigestion (H+P) was established and the optimal concentration and pH value for AR of bone specimens were determined. Second, the newly developed method was compared with existing AR methods (boiling in sodium citrate, hyaluronidase predigestion (H) and pepsin predigestion (P), with PBS only as the negative control) using two chromogenic detection systems (horseradish peroxidase (HRP) and alkaline phosphatase (AP)) to evaluate their efficacy in obtaining the best IHC results for bone samples. Considering the drawbacks of significant shrinking and detachment from slide for heat retrieval methods and the only moderate immunolabeling for H and P, H+P was the optimal AR method for IHC of bone specimens with the advantages of both good morphological preservation and strong immunoreactivity. Moreover, AP-mediated chromogenic detection was superior to HRP-labeled chromogenic detection due to significantly less nonspecific staining. In conclusion, we presented an effective and practical IHC protocol for bone specimens characterized by H+P predigestion combined with APmediated chromogenic detection. Finally, a detailed troubleshooting guide was provided for common mistakes that occur during IHC processing of the bone tissue samples.
Autor/es principal/es: Li, Shangfu
Liu, Bin
Tian, Ming
Zhang, Liangming
Tickner, Jennifer
Xu, Jiake
Rong, Limin
URI: http://hdl.handle.net/10201/88201
DOI: https://doi.org/10.14670/HH-30.331
Tipo de documento: info:eu-repo/semantics/article
Número páginas / Extensión: 13
Derechos: info:eu-repo/semantics/openAccess
Attribution-NonCommercial-NoDerivatives 4.0 International
Aparece en las colecciones:Vol.30, nº3 (2015)

Ficheros en este ítem:
Fichero Descripción TamañoFormato 
Li-30-331-343-2015.pdf10,53 MBAdobe PDFVista previa

Este ítem está sujeto a una licencia Creative Commons Licencia Creative Commons Creative Commons