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Flow cytometric analysis of Nkx2.2 expression in human embryonic kidney cells. 293-F cells (Life Technologies R790-07) were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574). Fixed cells were then stained with either PE Mouse IgG2b, κ Isotype Control (Cat. No. 555058, dashed line) or PE Mouse Anti-Nkx2.2 (solid line). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact 293-F cells. Flow cytometry was performed using a BD FACSCanto™ II Flow Cytometer System.
Flow cytometric analysis of Nkx2.2 expression in human embryonic kidney cells. 293-F cells (Life Technologies R790-07) were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574). Fixed cells were then stained with either PE Mouse IgG2b, κ Isotype Control (Cat. No. 555058, dashed line) or PE Mouse Anti-Nkx2.2 (solid line). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact 293-F cells. Flow cytometry was performed using a BD FACSCanto™ II Flow Cytometer System.
PE Mouse Anti-Nkx2.2 effectively stains 293-F cells when using either the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574) or BD Cytofix™ Fixation Buffer (Cat. No. 554655) with BD Phosflow™ Perm Buffer III (Cat. No. 558050).
The 74.5A5 monoclonal antibody specifically binds to Homeobox protein Nkx-2.2 (Nkx2.2), which is a transcription factor required for the differentiation of ventral neuronal populations in the central nervous system and endocrine cell populations in the pancreas and intestine. Within the pancreas, Nkx2.2 is expressed in many cell subsets and plays important roles in islet and β-cell differentiation and maturation. Mice with a homozygous disruption of the Nkx2-2 gene lack mature β cells and die shortly after birth with severe hyperglycemia. Similarly to the Nkx2.2-ablated mice, homozygous mutations in the human NKX2-2 gene have been described in diabetic patients and associated with the etiology of neonatal diabetes. The NKX2-2 gene is also a target of the Ewing sarcoma specific protein EWS-FLI. Nkx2.2 expression can be monitored in order to determine cell fate changes in cultures for the generation of human pluripotent stem cell-derived insulin-producing cells as well as oligodendrocyte progenitors.
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