NPY-mRFP Rat Basophilic Leukemia (RBL) Reporter: A Novel, Fast Reporter of Basophil/Mast Cell Degranulation.

Abstract

Humanized rat basophilic leukemia (RBL) reporter cell lines are increasingly used for the detection of allergen-specific IgE and other purposes, such as the detection of allergens and standardization of allergen preparations. Existing reporter systems have many strengths and advantages but can be expensive or require longer incubation times. The new NPY-mRFP reporter cell line addresses such problems, as it requires neither expensive substrates nor overnight incubation for detection of activation. The fusion of Neuropeptide Y (NPY) with monomeric Red Fluorescent Protein (mRFP) results in localization of the fluorescent protein in granules.AsNPY-mRFP is preformed in granules, the reporter systemactivation can be assessed using fluorescence measurements after as soon as 45-60min, as described in this chapter, without the need to add any substrates.