Quantifying IgA-Coated Bacteria by FCM

The proportion of IgA-coated bacteria (IgA-CB) in CC was determined by FCM as previously described [48 (link)]. The CC homogenate was stained with rabbit anti-rat Ig polyclonal antibody conjugated to fluorescein isothiocyanate (FITC) (Abcam, Cambridge, UK). Bacteria were gated in a FacsAria SORP sorter (BD Biosciences) after PI staining (Sigma-Aldrich) and according to their forward (FSC) and side scatter (SSC) characteristics. Analysis was performed in the FCU of the CCiTUB using the FlowJo v.10 software.

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Ruiz-Iglesias P., Estruel-Amades S., Massot-Cladera M., Franch À., Pérez-Cano F.J, & Castell M. (2022). Rat Mucosal Immunity following an Intensive Chronic Training and an Exhausting Exercise: Effect of Hesperidin Supplementation. Nutrients, 15(1), 133.

Publication 2022

FITC FacsAria SORP sorter

Anti antibody Bacteria Fluorescein Isothiocyanate Rabbit

Corresponding Organization : Instituto de Salud Carlos III

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Variable analysis

independent variables

Proportion of IgA-coated bacteria (IgA-CB) in CC

dependent variables

Proportion of IgA-coated bacteria (IgA-CB) in CC

control variables

Rabbit anti-rat Ig polyclonal antibody conjugated to fluorescein isothiocyanate (FITC)
Propidium iodide (PI) staining
Forward scatter (FSC) and side scatter (SSC) characteristics of bacteria
FlowJo v.10 software used for analysis

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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