The S25-763 monoclonal antibody specifically binds PTF1A (Pancreas specific transcription factor 1a). PTF1A is a transcription factor that contains a basic helix-loop-helix motif domain and, in combination with p75 and p64, forms the pancreas transcription factor complex. PTF1A plays a roles in pancreatic and cerebellar development. In early pancreatic development, pancreatic bud cells can either continue to pancreatic organogenesis or regress to a duodenal fate, and PTF1A plays a role in this differentiation. In late pancreatic development, the expression of PTF1A is restricted to acinar cells where it is involved in the maintenance of exocrine-specific gene expression. PTF1A expression is often evaluated to monitor pancreatic lineage differentiation from pluripotent stem cells. In addition, PTF1A is expressed during retinal development, and mutations in PTF1A cause cerebellar deficiencies. This mouse anti-human PTF1A monoclonal antibody, clone S25-763, has been demonstrated to cross-react with rat and mouse pancreatic acinar cells. Analysis of PTF1A expression in rat pancreatic acinar cells by western blot and flow cytometry. Left panel: Lysates of the AR42J cell line (ATCC CRL-1492) were electrophoresed (SDS-PAGE) in a 4-20 % Tris-Glycine polyacrylamide gel, transferred to PVDF membranes and then probed with 2 (lane 1), 1 (lane 2), 0.5 (lane 3), 0.25 (lane 4) mg/mL of Purified Mouse anti-PTF1A. Specific staining was detected with HRP Goat Anti-Mouse Ig (Cat. No. 554002). Right panel: AR42J cells were fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) and stained with matching concentrations of either Purified Mouse IgG1, κ Isotype Control (Cat. No. 554121, dashed line) or Purified Mouse anti-PTF1A monoclonal antibody (solid line). The second-step reagent was PE Goat Anti-Mouse Ig (550589). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact AR42J cells. Flow cytometry was performed using a BD LSRFortessa™ Flow Cytometer System. Immunohistochemical analysis of PTF1A expression by acinar cells within the human pancreas. Following antigen retrieval with BD Retrievagen A Buffer (Cat. No. 550524), the formalin-fixed paraffin-embedded sections were stained with either Purified Mouse IgG1 κ Isotype Control (Cat. No. 550878, left panel) or Purified Mouse anti-PTF1A antibody (right panel). A three-step staining procedure that employs Biotin Goat Anti-Mouse Ig (Cat. No. 550337), Streptavidin- HRP (Cat. No. 550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. The acinar cell nuclei and cytoplasm stained positively for the expression of PTF1A. Original magnification: 40x.