Quantifying Immunoglobulins in Mucosal Samples

The concentration of IgA and/or IgM in SMG, GW, CC, and feces were determined by a sandwich ELISA (Bethyl Laboratories Inc., Montgomery, TX, United States), as previously explained (36 (link)). Absorbance was quantified on a microplate photometer (Labsystem Multiskan, Helsinki, Finland) and data were interpolated by Ascent v.2.6 software (Thermo Fisher Scientific) according to the respective standard curves. The Ig content in SMG, CC and feces was normalized by the total protein concentration, which was determined using the Pierce-660 nm ready-to-use Protein Assay Reagent (Thermo Fisher Scientific) following the manufacturer’s instructions.

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Ruiz-Iglesias P., Massot-Cladera M., Rodríguez-Lagunas M.J., Franch À., Camps-Bossacoma M., Castell M, & Pérez-Cano F.J. (2022). A Cocoa Diet Can Partially Attenuate the Alterations in Microbiota and Mucosal Immunity Induced by a Single Session of Intensive Exercise in Rats. Frontiers in Nutrition, 9, 861533.

Publication 2022

sandwich ELISA Ascent v.2.6 software Pierce-660 nm ready-to-use Protein Assay Reagent

Assay Elisa Feces Protein

Corresponding Organization : Centro de Investigación Biomédica en Red

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Variable analysis

independent variables

Concentration of IgA and/or IgM

dependent variables

Absorbance measured on a microplate photometer
IgA and/or IgM content in submandibular gland (SMG), cervical cord (CC), and feces, normalized by total protein concentration

control variables

Total protein concentration, determined using the Pierce-660 nm ready-to-use Protein Assay Reagent

controls

No positive or negative controls were explicitly mentioned in the provided information.

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