ArticleMethods in molecular biology (Clifton, N.J.)2025
Use of Humanized NFAT-Dsred RBL Reporter for Detection of Allergen-Specific IgE Sensitization in Human Serum in 96-Well Microarray Format.
Article in Methods in molecular biology (Clifton, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
We have established a workflow allowing printing of allergens and control proteins into the wells of a MaxiSorp 96-well plate. Allergens are printed e.g., in a 4 × 4 pattern into each well of the plate. Prior to adding to the printed plates, a humanized rat basophil leukemia (RBL)-derived fluorescent reporter cell line (e.g., RBL NFAT DsRed) is sensitized overnight with the IgE-containing serum to be tested. Next, the humanized reporter cell line is added to the wells with the printed allergens and incubated for 24 h in a cell standard culture incubator. If the serum sample contains enough IgE specific for the printed allergens, NFAT-dependent signal transduction will occur, leading to synthesis of intracytosolic DsRed protein during an overnight incubation. Fluorescence can then be measured using a microplate reader using the well scan function.
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