Immunofluorescent Detection of Citrobacter in Mouse Colon

Mouse colons were embedded in OCT after euthanasia. The tissues were sectioned at 4 μm thickness, and fixed with 4% formaldehyde for 30 min at RT. After washing with PBS, the section was blocked with blocking buffer (Zymed Laboratories Inc., San Diego, USA) before incubated with anti-Citrobacter antibody (Abcam, ab37056) overnight at 4°C as previously described[41 (link)]. The section was then stained with Secondary Fluorescent Antibody. After washing in PBS, the tissues were counterstained with mounting medium containing DAPI.

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An J., Zhao X., Wang Y., Noriega J., Gewirtz A.T, & Zou J. (2021). Western-style diet impedes colonization and clearance of Citrobacter rodentium. PLoS Pathogens, 17(4), e1009497.

Publication 2021

ab37056

Anti antibody Buffer Citrobacter Colons Dapi Euthanasia Fluorescent antibody Formaldehyde Mouse Tissues

Corresponding Organization : Georgia State University

Other organizations : Huashan Hospital, Fudan University

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Variable analysis

independent variables

Tissue fixation with 4% formaldehyde for 30 min at RT

dependent variables

Expression of Citrobacter protein

control variables

Mouse colon tissue embedded in OCT
Tissue sectioned at 4 μm thickness
Blocking with blocking buffer before antibody incubation
Incubation with anti-Citrobacter antibody overnight at 4°C
Staining with Secondary Fluorescent Antibody
Counterstaining with mounting medium containing DAPI

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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