The tumors were fixed in 10% neutral buffered formalin and included in paraffin. 4μm-thick tissue sections obtained from paraffin embedded tissues were deparaffinized, rehydrated and stained with H&E to define tumor histotypes. Immunohistochemistry was performed using a polymer detection method. Briefly, the antigen retrieval was performed using Novocastra Epitope Retrieval Solution pH 9 in a PT Link Dako pre-treatment module at 98°C for 30 minutes. Subsequently, the sections were brought to room temperature and washed in PBS. After neutralization of the endogenous peroxidase with 3% H2O2 and Fc blocking by a specific protein block (Novocastra UK), the samples were incubated with primary antibodies. For rat anti-mouse monoclonal CD8a (4SM15) or CD4 (4SM95) 1:100 pH9 (eBioscience) antibodies, the staining was revealed using goat anti-rat IgG (H&L) specific secondary antibody 1:500 (Novex by Life Technologies) and AEC (3-Amino-9-ethylcarbazole) substrate-chromogen. Slides were analyzed under a Zeiss Axioscope A1 and microphotographs were collected using a Zeiss Axiocam 503 Color with the Zen 2.0 Software (Zeiss). Quantitative IHC data for CD8 and CD4 marker was calculated by counting the number of CD8+ or CD4+ cells in five fields at 40X magnification.
Cd4 4sm95
Manufactured by Thermo Fisher Scientific
Sourced in United States
The CD4 (4SM95) is a laboratory equipment designed for the detection and quantification of CD4 (Cluster of Differentiation 4) positive cells. It utilizes flow cytometry technology to analyze cell samples and provide accurate measurements of CD4 cell counts.
Automatically generated - may contain errors
Lab products found in correlation
Foxp3 fjk 16s, by Thermo Fisher Scientific (2 mentions)
Cd8 4sm15, by Thermo Fisher Scientific (2 mentions)
Axiocam 503 color, by Zeiss (1 mentions)
Specific protein block, by Leica (1 mentions)
Epitope retrieval solution ph 9, by Agilent Technologies (1 mentions)
Zen 2, by Zeiss (1 mentions)
Anti pd 1, by BioXCell (1 mentions)
Bkm120, by Novartis (1 mentions)
Mac 3, by BD (1 mentions)
Ra3 6b2, by BD (1 mentions)
Rmab005, by Diagnostic BioSystems (1 mentions)
4 protocols using cd4 4sm95
1
Immunohistochemical Analysis of Tumor-Infiltrating T Cells
2
Pharmacokinetics and Immune Modulation
3
Artery Graft Histological Analysis
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Artery grafts were perfusion fixed with 4% paraformaldehyde. For histological examination of medial injury, 2 cross-sections per artery that were at least 100 μm apart were H&E stained and the average thickness of the media measured in Image J (National Institutes of Health). The medial thickness was normalized to arterial radius.
Immunohistochemistry was performed using antibodies toward myeloperoxidase (Abcam, Cambridge, United Kingdom) to visualize neutrophils, CD4 (4SM95; eBioscience, Waltham, Massachusetts), CD8 (4SM15, eBioscience), Foxp3 (FJK-16S, eBioscience), and Mac-3 (BD Biosciences, Franklin Lakes, New Jersey) to visualize macrophages as described previously. 29 Staining was visualized by aminoethyl carbazole chromagen (red; Vector Laboratories, Burlingame, CA) and counterstained with hematoxylin (blue). For quantification, the total number of cells in each cross-section was manually counted in a blinded manner and reported per mm 2 of media or adventitia. Two cross-sections that were at least 100 μm apart were evaluated for each artery and an average value from the cross-sections calculated and used as a single data point.
Immunohistochemistry was performed using antibodies toward myeloperoxidase (Abcam, Cambridge, United Kingdom) to visualize neutrophils, CD4 (4SM95; eBioscience, Waltham, Massachusetts), CD8 (4SM15, eBioscience), Foxp3 (FJK-16S, eBioscience), and Mac-3 (BD Biosciences, Franklin Lakes, New Jersey) to visualize macrophages as described previously. 29 Staining was visualized by aminoethyl carbazole chromagen (red; Vector Laboratories, Burlingame, CA) and counterstained with hematoxylin (blue). For quantification, the total number of cells in each cross-section was manually counted in a blinded manner and reported per mm 2 of media or adventitia. Two cross-sections that were at least 100 μm apart were evaluated for each artery and an average value from the cross-sections calculated and used as a single data point.
4
Immunohistochemical Profiling of T-cells and Macrophages
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IHC staining was performed with BondMax with Refine HRP-Kit DS9800 (Leica Biosystems, Muttenz, Switzerland) in accordance with the manufacturer's guidelines. Primary antibodies were directed against CD3 (RMAB005; Diagnostic Biosystems, Pleasanton, CA, USA), CD4 (4SM95; eBioscience, San Diego, CA, USA), F4/80 (T-1006; BMA Biomedicals, Augst, Switzerland), and rat CD45 (B220) (RA3-6B2; BD Pharmingen, Allschwil, Switzerland). The number of positive cells in the perivascular and
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