Mouse aortas were prepared and embedded in Tissue-Tek (Sakura Finetek), and serial 10-μm-thick frozen tissue sections were prepared. Immunostainings were performed using following marker antibodies: anti-mouse CD3e (145–2C11, BD; 1:100 dilution), anti-mouse Foxp3 (ab75763, Abcam; 1:100 dilution) and DAPI for DNA. Secondary antibodies were used as previously described7 (link). For negative controls, stainings were performed without primary antibodies or isotype controls. Stained sections were analyzed using a Leica SP8 ×3 microscope (Leica microsystems, Germany). Images were acquired with identical microscope settings using sequential channel acquisitions to avoid cross-talk between fluorophores. All images were prepared as TIF files by Fiji (ImageJ, National Institutes of Health) or Leica LAS-X (V3.5) software and exported into Adobe Illustrator CS6 for figure arrangements.
Anti mouse cd3e 145 2c11
Manufactured by BD
Sourced in United Kingdom
Anti-mouse CD3e (145–2C11) is a monoclonal antibody that recognizes the CD3 epsilon chain, a component of the T cell receptor complex in mice. It is commonly used in flow cytometry and other immunological applications to identify and study T cells.
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3 protocols using anti mouse cd3e 145 2c11
1
Immunostaining of Aortic Tissue Sections
2
Immunostaining of Aortic Tissue Sections
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Mouse aortas were prepared and embedded in Tissue-Tek (Sakura Finetek), and serial 10-μm-thick frozen tissue sections were prepared. Immunostainings were performed using following marker antibodies: anti-mouse CD3e (145–2C11, BD; 1:100 dilution), anti-mouse Foxp3 (ab75763, Abcam; 1:100 dilution) and DAPI for DNA. Secondary antibodies were used as previously described7 (link). For negative controls, stainings were performed without primary antibodies or isotype controls. Stained sections were analyzed using a Leica SP8 ×3 microscope (Leica microsystems, Germany). Images were acquired with identical microscope settings using sequential channel acquisitions to avoid cross-talk between fluorophores. All images were prepared as TIF files by Fiji (ImageJ, National Institutes of Health) or Leica LAS-X (V3.5) software and exported into Adobe Illustrator CS6 for figure arrangements.
3
Characterization of ASKP1240 Binding
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Peripheral blood samples from mice (B6C3F1), rats (Sprague–Dawley), rabbits (Japanese White), cynomolgus monkeys and humans were incubated with FITC-labeled ASKP1240. The erythrocytes/platelets, lymphocyte, monocytes and granulocytes were identified by their light scatter characteristics and lineage-specific mAbs (anti-mouse CD3e, 145-2C11, BD; anti-CD45R/B220, RA3-6B2, BD; anti-mouse CD41, MWReg30, BD; anti-rat CD3, G4.18, BD; anti-rat CD45RA, OX-33, BD; anti-rat CD42d, RPM.4, BD; anti-rabbit T lymphocyte, KEN-5, AbD Serotec, Kidlington, UK; anti-rabbit CD41, P2, Beckman Coulter, Brea, CA; anti-monkey CD3, SP34 BD; anti-monkey CD20, H29, Beckman Coulter; anti-monkey CD4a1, HIP8, BD; anti-human CD3, UCHT1, Beckman Coulter; anti-human CD19, J4.119, Beckman Coulter; anti-human CD41, P2, BD). The fluorescent activity of each cell population was determined by flow cytometry.
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