The FFPE tissue samples were cut into 4 mm thin sections. Slides were incubated for 10 min in 0.3% hydrogen peroxide. Subsequently, sections were incubated overnight at 4°C with the primary antibody. Then, slides were incubated with a secondary antibody [either EnVision + System-HRP Labeled Polymer anti-rabbit or anti-mouse (Dako, Agilent, Santa Clara, United States)]. The average density (cells/mm2) of each lymphocyte subset was calculated from five 1 mm2 regions within each tumor to calculate its average density of positive cells (Reuben et al., 2017 (link)). IPMN was confirmed in all tissue samples by H&E staining, and the grade of dysplasia was evaluated by two experienced pathologists (ZJY and HX).
Envision system hrp labeled polymer anti rabbit or anti mouse
Manufactured by Agilent Technologies
Sourced in United States
The EnVision + System-HRP Labeled Polymer anti-rabbit or anti-mouse is a laboratory equipment product from Agilent Technologies. It is designed to be used as a detection reagent in immunohistochemistry and other related applications. The product utilizes a polymer-based detection system that is labeled with horseradish peroxidase (HRP) for the detection of rabbit or mouse primary antibodies.
Automatically generated - may contain errors
4 protocols using envision system hrp labeled polymer anti rabbit or anti mouse
1
Immunohistochemical Quantification of Lymphocyte Subsets in IPMN
2
Immunohistochemical Analysis of Xenograft Tumors
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For IHC testing, the xenograft tumor specimens were stained with either anti-rabbit polyclonal β-catenin or anti-mouse USP4 (Santacruz, USA). After deparaffinization and rehydration, formalin-fixed, paraffin-embedded xenograft tumor sections were incubated in PBS containing 0.3% H 2 O 2 for 10 min at room temperature followed by an antigen retrieval step carried out by boiling the slides in 0.01 M citrate buffer for 5 min. The sections were then washed in 50 mM Tris-HCl pH 7.6 and 0.05 % Tween20 for 2 min. To block nonspecific binding, all the sections were treated with PBS containing 2% BSA and 2% goat serum for 30 min at room temperature. The slides were then incubated with a primary antibody (rabbit polyclonal anti-β-catenin antibody or mouse anti-USP4 antibody) in the blocking solution at 4 °C overnight. The reaction was stopped by rinsing the section with PBS. The slides were then incubated with a Dako EnVision+ System HRP-labeled polymer anti-rabbit or anti-mouse (Agilent, USA) and treated with diaminobenzidine substrate to visualize the positive cells. Finally, the sections were counterstained with hematoxylin before being mounted for light microscopy examination.
3
Immunohistochemical Analysis of Tumor Samples
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Samples were fixed and processed as described in [19 (link)]. The following primary antibodies were used: mouse monoclonal anti-CD3 (clone SP7, 1:70, Leica, #565-LCE) and anti-FOXP3 (1:200, Abcam, ab22510), rat monoclonal anti-Ly6G (clone 1A8, 1:400, Cederlane, #ABF118UD), rabbit polyclonal anti-IBA1 (1:300, Wako, #019-19741), and mouse monoclonal anti-TfR (1:800 #M36008 Invitrogen). Detection was done using EnVision+ System-HRP Labeled Polymer anti-mouse or anti-rabbit (Dako) or using Rat-on-Mouse HRP-Polymer (Biocare Medical) followed by DAB as chromogen. Hematoxylin counterstaining was performed on all sections.
Positive cells count was carried out with Aperio Imagescope on digitalized sections acquired using the Aperio CS2 digital scanner and ScanScope software. The analysis was perform using IHC nuclear Image Analysis and Positive Pixel Count v9 9.0 Algorithm (Imagescope, Leica Biosystem). The whole tumor area was considered for the analysis and necrotic areas were excluded.
Positive cells count was carried out with Aperio Imagescope on digitalized sections acquired using the Aperio CS2 digital scanner and ScanScope software. The analysis was perform using IHC nuclear Image Analysis and Positive Pixel Count v9 9.0 Algorithm (Imagescope, Leica Biosystem). The whole tumor area was considered for the analysis and necrotic areas were excluded.
4
Histological Analysis of SARS-CoV-2 Spike Protein
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Lung tissues were fixed with 10% neutral buffered formalin and embedded in paraffin. 2 μm tissue sections were prepared and stained with Hematoxylin and eosin (H&E). For immunohistochemical staining, 2 μm thickness sections were immersed in citrate buffer (pH 6.0) and heated for 20 min with a pressure cooker. Endogenous peroxidase was inactivated by immersion in 3% H2O2 in PBS. After treatment with 5% BSA in PBS for 30 min at room temperature, the sections were incubated with mouse anti-Spike protein antibody (1:300, clone 1A9, Genetex, Irvine, CA, USA). For secondary antibody, EnVision+ system-HRP-labeled polymer anti-mouse or anti-rabbit (Dako, Carpinteria, CA, USA) were used. Positive signals were then visualized by peroxidase–diaminobenzidine reaction and sections were counterstained with hematoxylin.
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