Microslide tissue sections were deparaffinized with xylene, hydrated using a diluted alcohol series, and immersed in 0.3% H2O2 in methanol to extinguish endogenous peroxidase activity. Sections were then microwaved for 15 minutes in 10 mM citrate buffer (pH 6.0) for antigen retrieval. Each section was blocked with 4% bovine serum albumin in phosphate buffered saline with 0.1% Tween 20 (PBST) for 30 minutes to reduce non-specific staining. Sections were incubated with anti–PD-L1 (1:100, Cell Marque, Rocklin, CA, USA) or anti–PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies in PBST containing 3 mg/mL goat globulin (Sigma-Aldrich, St. Louis, MO, USA) for 60 minutes at room temperature, followed by three successive washes with a buffer. The sections were then incubated with an antimouse/rabbit antibody (Envision plus, Dako, Carpinteria, CA, USA) for 30 minutes at room temperature. The chromogen used was 3,3'-diaminobenzidine (Dako). The sections were counterstained with Meyer’s hematoxylin. For positive controls, sections of human placenta and tonsil tissue were included in each staining run. Omission of the primary antibody for placenta and tonsil tissue sections was used as a negative control.
Anti pd 1
Manufactured by Roche
Sourced in United States, United Kingdom
Anti-PD-1 is a laboratory equipment product. It is used to detect and measure the presence of PD-1 (Programmed Cell Death Protein 1) in biological samples.
Automatically generated - may contain errors
2 protocols using anti pd 1
1
Immunohistochemical Detection of PD-L1 and PD-1
2
Immunohistochemical Analysis of Tumor-Infiltrating Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies. All slides were evaluated and scored under light microscopy, analyzed and photographed by using Olympus cellSens software (version 1.4). Tumor-infiltrating inflammatory cells (TIICs) were identified based on morphology in hematoxylin–eosin (H&E)-stained sections. Immunohistochemical staining was measured by semiquantitative assessment of positive staining in TIICs. For VISTA, CD33, and PD-1 antibodies, the percentage of TIICs showing positive membrane staining was determined. The intensity of staining was determined on a scale of 0–3 (0 = < 5%, 1 = 5–20%, 2 = 20–50%, and 3 = > 50% of TIICs). Cases with a score ≥ 1 were considered positive. Scores of 2 or 3 were considered to represent high expression of VISTA, CD33, and PD-1. The numbers of VISTA-, PD-1-, and CD33-positive TIICs were counted in five representative high-powered fields (200X magnification) and averaged. All melanoma tissues evaluated in the study are the primary tumor tissues from the primary cutaneous melanoma tissues. All samples were evaluated independently by two investigators (LWJ, CJW).
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!