Immunohistochemistry (IHC) was performed automatically on a Ventana BenchMark Ultra instrument (Ventana Medical System, Tucson, Arizona, USA). The primary antibody used was c-FOS (E8): sc-166940 (Santa Cruz Biotechnology, Dallas, Texas, USA) diluted 1:600. After deparaffination and rehydration of the sections, antigen retrieval was done using EDTA buffer at 100 °C for 30 min. Slides were treated with inhibitors to endogenous peroxidase followed to incubation with primary antibody in 30 min at 37 °C. Amplification was done using Optiview system (Ventana Medical System, Tucson, Arizona, USA). Visualization was done using Diaminobenzidine (DAB) and Hematoxylin was used as counterstain. Human term placenta was used as positive control.
Optiview system
Manufactured by Roche
Sourced in United States
The Optiview system is a laboratory equipment product designed for automated optical analysis. It provides precise and reliable measurements for a variety of applications. The core function of the Optiview system is to perform optical analysis of samples, enabling researchers and analysts to obtain accurate data and insights.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using optiview system
1
Automated c-FOS Immunohistochemistry Protocol
2
IHC Staining for BRAF V600E
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The CRC tumor microarray was cut at 4-mm thick sections for IHC studies. Deparaffinized sections were stained on a Ventana (Tucson, AZ) Benchmark XT automated immunostainer with the use of a 32-minute CC1 heat-induced antigen retrieval protocol. Incubation with VE-1 mouse monoclonal primary antibody (Spring Biosciences, Pleasanton, CA) directed against B-Raf V600E protein was for 16 minutes at 37 C. Antibody was detected with the Ventana Optiview system with the use of diaminobenzidine as a chromogen. Previous experience with this antibody suggested that homogeneous cytoplasmic staining of carcinoma cells indicates the presence of B-Raf V600E protein.
Nonspecific nuclear staining, readily distinguished from cytoplasmic staining, was encountered in both benign and malignant colorectal epithelium in some cases. Blinded IHC scoring was performed by two pathologists (R.W. and Brandon Sheffield) and a histotechnologist (Deanna Johnson) over a multiheaded microscope. Tumor microarray cases scored as indeterminate were retested by IHC on the whole section from which the corresponding tumor microarray core was taken and then rescored.
Nonspecific nuclear staining, readily distinguished from cytoplasmic staining, was encountered in both benign and malignant colorectal epithelium in some cases. Blinded IHC scoring was performed by two pathologists (R.W. and Brandon Sheffield) and a histotechnologist (Deanna Johnson) over a multiheaded microscope. Tumor microarray cases scored as indeterminate were retested by IHC on the whole section from which the corresponding tumor microarray core was taken and then rescored.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!