Immunohistochemical Analysis of Prostate Cancer

Immunohistochemistry (IHC) was performed on tissue arrays from PCa (CA3, SuperBioChips Tissue Array, Tema Ricerca, Italy) as previously described [35 (link)]. The primary antibodies used were as follows: anti-p65 (RB-1638-R7, ready to use, NeoMarker, Thermo Fisher Scientific, Fremont, CA, USA); anti-Shh (sc-1194 N19) (1:50, Santa Cruz Biotechnology); anti-GLI1 (sc-20687 H-300) (1:50, Santa Cruz Biotechnology, Dallas, Texas, USA). Immunoreactions were visualized using the Avidin/Biotinylated enzyme Complex (ABC) (VECTASTAIN Elite ABC System, Burlingame, CA, USA) and sections were counterstained using Mayer’s hematoxylin (Emallume Carazzi Bio-Optica, Milano, Italy). Histological images were acquired using a Nikon Eclipse E200 microscope equipped with a Leica DFC310 FX Digital Camera. The percentage of positive area was quantified using the QuPath Software [36 (link)].

Free full text: Click here

Vecchiotti D., Verzella D., Di Vito Nolfi M., D’Andrea D., Flati I., Di Francesco B., Cornice J., Alesse E., Capece D, & Zazzeroni F. (2022). Elevated NF-κB/SHh/GLI1 Signature Denotes a Worse Prognosis and Represent a Novel Potential Therapeutic Target in Advanced Prostate Cancer. Cells, 11(13), 2118.

Publication 2022

NeoMarker Elite ABC System Eclipse E200 microscope DFC310 FX Digital Camera

Antibodies Avidin Camera Enzyme complex Hematoxylin Immunohistochemistry Microscope Tissue

Corresponding Organization : University of L'Aquila

Other organizations : Nottingham Trent University

Top 5 similar protocols

Variable analysis

independent variables

Primary antibodies used: anti-p65, anti-Shh, anti-GLI1

dependent variables

Percentage of positive area quantified using QuPath Software

control variables

Tissue arrays from PCa (CA3, SuperBioChips Tissue Array, Tema Ricerca, Italy)
Immunohistochemistry (IHC) protocol as previously described [35]
Avidin/Biotinylated enzyme Complex (ABC) (VECTASTAIN Elite ABC System, Burlingame, CA, USA) for visualizing immunoreactions
Sections counterstained using Mayer's hematoxylin (Emallume Carazzi Bio-Optica, Milano, Italy)
Histological images acquired using a Nikon Eclipse E200 microscope equipped with a Leica DFC310 FX Digital Camera

controls

No positive or negative controls were explicitly mentioned in the provided information.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!