Immunofluorescence Analysis of Murine IDO1

Immunofluorescence on frozen sections was performed as previously described (21 (link)). The following antibodies were used: polyclonal goat anti-mouse IDO1 (I-17 Cat # sc-25121, or M-20 Cat # sc-25123, Santa Cruz Biotechnology, Santa Cruz, CA); FITC conjugated monoclonal hamster anti CD11c (clone N418, Cat # 117306, BioLegend, San Diego, CA); monoclonal rabbit anti E-Cadherin (clone 24E10, Cat #3195, Cell Signaling, Boston, MA);FITC conjugated monoclonal mouse anti MPO (clone 2D4, Cat #ab90812, Abcam, Cambridge, MA); Alexa Fluor 647 conjugated (far red) monoclonal rat anti IFNγ (clone XMG1.2, Cat #505816, BioLegend). Staining was visualized using an Olympus Fluoview scanning confocal microscope (Olympus, Center Valley, PA).

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El-Zaatari M., Chang Y.M., Zhang M., Franz M., Shreiner A., McDermott A.J., van der Sluijs K.F., Lutter R., Grasberger H., Kamada N., Young V.B., Huffnagle G.B, & Kao J.Y. (2014). Tryptophan Catabolism Restricts IFN-γ-expressing Neutrophils and Clostridium difficile Immunopathology. Journal of immunology (Baltimore, Md. : 1950), 193(2), 807-816.

Publication 2014

Fluoview scanning confocal microscope

Alexa fluor 647 Antibodies Cadherin Clone Confocal microscope Fitc Frozen sections Goat Hamster Ifnγ Immunofluorescence Mouse Rabbit

Corresponding Organization : University of Michigan–Ann Arbor

Other organizations : Academic Medical Center

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Variable analysis

independent variables

Polyclonal goat anti-mouse IDO1 (I-17 Cat # sc-25121, or M-20 Cat # sc-25123, Santa Cruz Biotechnology, Santa Cruz, CA)
FITC conjugated monoclonal hamster anti CD11c (clone N418, Cat # 117306, BioLegend, San Diego, CA)
Monoclonal rabbit anti E-Cadherin (clone 24E10, Cat #3195, Cell Signaling, Boston, MA)
FITC conjugated monoclonal mouse anti MPO (clone 2D4, Cat #ab90812, Abcam, Cambridge, MA)
Alexa Fluor 647 conjugated (far red) monoclonal rat anti IFNγ (clone XMG1.2, Cat #505816, BioLegend)

dependent variables

Immunofluorescence staining pattern

control variables

Frozen sections
Olympus Fluoview scanning confocal microscope (Olympus, Center Valley, PA)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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