Paraffin-embedded sections of human colon from CD patients and controls were cut (5 μm) and deparaffinized. For immunohistochemistry, sections were microwaved for 2 × 5-min cycles in 10 mM citrate buffer, and processed by ABC method according to the manufacturer’s protocol (Vectastain ABC kit; Vector Laboratories, Burlingname, CA, USA). Sections were incubated with rabbit anti-CRTH2 (1:200; Acris Antibodies, Herford, Germany) 11 (link), visualized with 3-3′-diaminobenzidine (DAB) and counterstained with hematoxylin. CD4+ T cells were stained with a monoclonal mouse anti-human CD4 (clone 4B12; dilution 1:20; Labvision, Fremont, USA) as recommended by the suppliers. Images were taken with a high resolution digital camera (Olympus DP 50) and analyzed by Cell^A imaging software (Olympus, Vienna, Austria). Only contrast and brightness of images were adjusted. Sirius Red (Direct Red 80®, Sigma) was used to stain eosinophils in deparaffinized sections.
Monoclonal mouse anti human cd4 clone 4b12
Manufactured by Lab Vision
Sourced in United States, Denmark
Monoclonal mouse anti-human CD4 (clone 4B12) is a laboratory reagent used for the identification and enumeration of CD4-positive cells in human biological samples. It is a primary antibody that specifically binds to the CD4 cell surface receptor.
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2 protocols using monoclonal mouse anti human cd4 clone 4b12
1
Immunohistochemical Analysis of CRTH2 and CD4+ T Cells in Colon Tissues
2
Immune Profiling of Gastric Mucosa
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Formalin‐fixed paraffin‐embedded (FFPE) biopsy specimens were derived from the files of the Institute of Pathology of the Medical University of Graz (supplementary material, Table S1). Only cases with paired duodenal, gastric antral and gastric corpus specimens were included in the study. H. pylori carriage was determined by Warthin–Starry staining 19 and/or immunohistochemistry with an anti‐H. pylori antibody (clone SP48; Ventana, Tucson, AZ, USA). The following entities were used: healthy corpus (n = 24), corpus biopsies from corpus‐dominant LyG with proven absence of CeD (denoted LyG, n = 25), and corpus biopsies of H. pylori gastritis (denoted HpG, n = 25). Metadata and analyses performed on specimens are provided in supplementary material, Table S1. Sections from FFPE tissue specimens were stained with monoclonal mouse anti‐human CD8 (clone C8/144B; dilution 1:30; Dako Glostrup, Denmark), monoclonal mouse anti‐human CD4 (clone 4B12; dilution 1:20; Labvision, Fremont, CA, USA) and MICA/B (clone F‐6; dilution 1:200; Santa Cruz Biotechnology, Dallas, TX, USA) antibodies, according to the supplier's recommendations.
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