Phospho-PKR Immunohistochemistry in Ovarian Cancer

The tissue array consists of matched recurrent/resistant and primary/naive from 14 ovarian cancer patients (39 (link)). Slides deparaffinization, antigen retrieving, and blocking were performed as we have described (40 (link)). The arrays were stained with anti-phospho- PKR T446 antibody (ab32036, 1:200 dilutions, Abcam) by using a VECTASTAIN Elite ABC-Peroxidase Kits following the manufacturer’s instructions (PK-6101, VECTOR LAB INC). Mouse tumors from SKOV3 xenografts (control and PKR knockout) were used to validate the antibody. Cleaved caspase 3 IHC staining (1:100 dilutions) was also used to detect cell death in tumors. Cell nuclei were stained with hematoxylin. Ventana iScan HT (Roche) was used for slide scanning with a 20X objective lens. Scoring and statistical analysis were done as we previously described (40 (link)).

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Yin L., Zeng Y., Zeng R., Chen Y., Wang T.L., Rodabaugh K.J., Yu F., Natarajan A., Karpf A.R, & Dong J. (2021). Protein Kinase RNA-activated Controls Mitotic Progression and Determines Paclitaxel Chemosensitivity through B-cell lymphoma 2 in Ovarian Cancer. Oncogene, 40(50), 6772-6785.

Publication 2021

Ventana iScan HT

Anti antibody Antibody Antigen Caspase 3 Cell death Cell nuclei Dilutions Lens Mouse Ovarian primary cancer Patients Peroxidase Tissue Tumors Vector Xenografts

Corresponding Organization :

Other organizations : University of Nebraska Medical Center, Johns Hopkins University, Sidney Kimmel Comprehensive Cancer Center

Top 5 similar protocols

Variable analysis

independent variables

PKR knockout in SKOV3 xenograft tumors

dependent variables

Phospho-PKR T446 expression measured by IHC
Cleaved caspase 3 expression measured by IHC

control variables

SKOV3 xenograft tumors (control)
Slide deparaffinization, antigen retrieving, and blocking procedures
IHC staining protocol using VECTASTAIN Elite ABC-Peroxidase Kits
Slide scanning using Ventana iScan HT (Roche) with 20X objective lens
Scoring and statistical analysis methods

positive controls

Mouse tumors from SKOV3 xenografts (control)

negative controls

Mouse tumors from SKOV3 xenografts (PKR knockout)

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!