Generation of Monoclonal Antibodies Against Mouse Neurogenin 3: A New Immunocytochemical Tool to Study the Pancreatic Endocrine Progenitor Cell

The induction of pancreatic endocrine differentiation from undifferentiated precursor cells is a multi-step process regulated by the expression of several transcription factors. At E9.5 expression of homeodomain protein Pdx-1 defines the early pancreatic epithelium. Later, expression of the basic helix-loop-helix factor, Neurogenin 3 (Ngn3) marks the initiation of the endocrine lineage from the pancreatic precursor cells. A full understanding of these processes is essential in order to control development of insulin secreting beta-cells from embryonic stem cells in vitro. Since the expression of ngn3 is a key step, the aim of this work was to develop monoclonal antibodies for immunohistochemical (IHC) detection of Ngn3. All mice (RBF-strain) immunized with recombinant GST-mNGN3 fusion protein responded with a high antibody titer. The sera were further screened by IHC on paraffin sections of fetal (E13) mouse pancreas, and two hybridomas subsequently derived from one selected mouse produced antibodies with...