Endoscopically or surgically collected biopsy samples were submitted for histopathological examination. Tissues were fixed in 10% buffered formalin and embedded in paraffin. Four micrometre‐thick sections were deparaffinised and stained with haematoxylin and eosin. Histopathological specimens were subjected to immunohistochemistry as needed, according to the discretion of the pathologist. The following primary antibodies were used for immunohistochemistry: rabbit anti‐CD3 antibody (ready to use; Dako, Tokyo, Japan), rabbit anti‐CD20 antibody (1:100; Biocare Medical, Pacheco, CA, USA), mouse anti‐CD79 antibody (1:50; clone HM57; GeneTex, Irvine, CA, USA), rabbit anti‐PAX5 antibody (1:50; Thermo Fisher Scientific, Waltham, MA, USA), rabbit anti‐lambda light chain antibody (1:5000; Dako, Tokyo, Japan) and rabbit anti‐kappa light chain antibody (1:1000; Dako, Tokyo, Japan). Horseradish peroxidase (Envision+ System; Dako, Tokyo, Japan) was used as secondary antibody and visualised with DAB chromogen.
Rabbit anti cd3 antibody
Manufactured by Agilent Technologies
Sourced in Japan
Rabbit anti-CD3 antibody is a laboratory reagent used in various immunological and cell biology applications. It binds specifically to the CD3 complex, a key component of the T cell receptor, and can be used to identify and study T cells.
Automatically generated - may contain errors
Lab products found in correlation
Dako envision kit, by Agilent Technologies (2 mentions)
Decloaking chamber, by Biocare Medical (2 mentions)
Monoclonal anti murine cd138 antibody, by BD (2 mentions)
Envision system, by Agilent Technologies (1 mentions)
Axiophot photomicroscope, by Zeiss (1 mentions)
Rat anti f4 80 antibody, by Bio-Rad (1 mentions)
Rat anti cd31 antibody, by Dianova (1 mentions)
Histofine reagent, by Nichirei Biosciences (1 mentions)
Guinea pig anti nephrin antibody, by Progen Biotechnik (1 mentions)
Prism 6, by GraphPad (1 mentions)
Rabbit anti cleaved caspase 3 antibody, by Cell Signaling Technology (1 mentions)
Metamorph, by Molecular Devices (1 mentions)
5 protocols using rabbit anti cd3 antibody
1
Immunohistochemistry for Tissue Biopsies
2
Histopathological and Immunohistochemical Analysis of Kidneys
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Kidneys were immersed in 10% formalin and embedded in paraffin. Sections (4 μm thick) were stained with hematoxylin/eosin or Masson's trichrome and processed for histopathology or immunohistochemistry. For immunohistochemistry, paraffin-embedded sections were stained with rabbit anti-nytrotyrosine primary antibody (Upstate, number 06284, 1 : 200), rabbit anti-CD3 antibody (DAKO, number A0452), and rat anti-F4/80 antibody (AbD Serotec, number MCA497R) then the staining was revealed with Histofine reagent (Nichirei Biosciences) and slides were counterstained with hematoxylin. For immunofluorescence, paraffin-embedded sections were stained with a guinea pig anti-nephrin antibody (Progen) and a rat anti-CD31 antibody (Dianova, number SZ31) then the stainings were revealed with a goat anti-guinea pig alexa568-coupled antibody and a donkey anti-rat alexa488-coupled antibody (Invitrogen) and nuclei were counterstained with DAPI. Photomicrographs were taken with an Axiophot Zeiss photomicroscope. Staining surface quantifications were performed with a macro designed on ImageJ.
3
Murine Liver Immunohistochemistry Protocol
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Murine liver sections (♀, balb/c, Taconic) were incubated in primary antibody solutions (rabbit anti-Alexa488 antibody (Molecular Probes) at 0.5 μg ml−1; rabbit anti-CD3 antibody (DAKO) at 1.2 μg ml−1; rabbit anti-Cleaved Caspase-3 antibody (Cell Signalling) at 0.1 μg ml−1; or TdT-biotin-dUTP (Roche) for TUNEL staining. Representative snapshots were taken from the scanned images and analysed for signal intensity or counts, normalized to tissue area, using image analysis software Metamorph (Molecular Devices, PA). Statistics (Student's t-tests) were performed and graphs were made using Prism6 (GraphPad, LaJolla, CA).
4
Immunohistochemical Analysis of CD3 and CD138
5
Immunohistochemical Analysis of CD3 and CD138
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4 μm thick formalin-fixed, paraffin-embedded tissue sections were examined. Slides were pre-treated with 10 mM citrate, pH 6.0 (Zymed, South San Francisco, CA) for CD3 or 1.0 mM EDTA pH 8.0 (Zymed) for CD138 in a pressure cooker (Decloaking Chamber, BioCare Medical, Walnut Creek, CA). Pre-diluted polyclonal rabbit anti-CD3 antibody (DAKO, Carpintera, CA) or monoclonal anti-murine CD138 antibody (Pharmingen, San Diego, CA) diluted 1:250 was applied for 1 hour at room temperature. Rabbit anti-rat immunoglobulin antibody at 1:7500 dilution and DAKO Envision kits (DAKO) were used for detection.
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