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Anti foxp3

Manufactured by Agilent Technologies
Sourced in Denmark

Anti-Foxp3 is a laboratory equipment product designed for the detection and analysis of the Foxp3 transcription factor. Foxp3 is a key regulator of regulatory T cells (Tregs) and is widely used as a marker for their identification. The Anti-Foxp3 product provides researchers with a tool to accurately measure and study Foxp3 expression, which is crucial for understanding the role of Tregs in various biological and pathological processes.

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2 protocols using anti foxp3

1

Immunofluorescence Analysis of CD3 and Foxp3

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Sections of formalin-fixed paraffin-embedded hearts and frozen spleens were used for detection of CD3 and Foxp3 expression by immunofluorescence. First, paraffin-embedded sections were deparaffinized and submitted to a heat-induced antigen retrieval step by incubation in citrate buffer (pH 6.0). Then, sections were incubated overnight with the following primary antibodies: anti-CD3 (1:400; BD Biosciences) and anti-Foxp3 (1:400; Dako, Glostrup, Denmark). On the following day, secondary antibodies anti-goat IgG Alexa Fluor 488 (1:600; Molecular Probes) or anti-rabbit IgG Alexa Fluor 568 conjugated (1:100; Molecular Probes), diluted in 1% BSA in PBS, were added. Nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI; VectaShield Hard Set mounting medium with DAPI H-1500; Vector Laboratories, Burlingame, CA). Images were analyzed using a confocal laser scanning microscope A1R (Nikon, Tokyo, Japan) and Image-Pro Plus version 7.01 (Media Cybernetics, Rockville, MD). Quantifications of CD3+/Foxp3+ cells percental area were performed in 10 fields randomly captured under x400 magnification, using Image-Pro Plus v.7.0.
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2

Immunohistochemical Analysis of Lung Markers

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Paraffin embedded lungs were sectioned, deparaffinized, hydrated in deionized water, and treated with Target Retrieval Solution (DAKO, Carpinteria, CA) at either pH 9 (anti-CD3, anti-Foxp3, anti-Ly6C/Ly6G) or pH 6 (anti-F4/80). All slides were then stained using a DAKO Autostainer Plus (DAKO, Carpinteria, CA) with the standard labeled streptavidin-biotin protocol. Lung sections were stained at room temperature for 60 min with polyclonal rabbit anti-CD3 (DAKO; 1:100), anti-F4/80 (AbD Serotec; clone A3-1; 1:250), anti-Foxp3 (eBioscience, clone FJK-16s; 1:25) or anti-Ly6C/Ly6G (clone NIMP-R14; 1:250). Negative controls were performed with omission of the primary antibody. Slides were then developed with Tertiary Streptavidin-HRP (DAKO) and counterstained with hematoxylin.
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