Rabbit anti pax6 prb 278p

Individual LEC cultures from three healthy and three diabetic corneas and corneoscleral rims were fixed in p-formaldehyde (1%, 10 min at 4 °C) followed by permeabilization in 0.25% Triton X-100 for 5 min at room temperature or in ice-cold methanol (10 min at 4 °C). The expression of putative LESC markers ΔNp63α and PAX6, keratins K12, K15, and K17, and ATP-binding cassette transporter ABCG2 was examined with immunostaining. Additionally, cultures were stained for differentiated corneal epithelial marker K12. The following primary antibodies were used: goat anti-ΔNp63α (sc-8609), goat anti-K12 (sc-17099), mouse anti-K15 (sc-47697), mouse anti-K17 (sc-58726; all from Santa Cruz Biotechnology, Dallas, TX), mouse anti-ABCG2 (MAB4155, Millipore, Billerica, MA), and rabbit anti-PAX6 (PRB-278P, BioLegend, San Diego, CA). Secondary antibodies conjugated either with fluorescein isothiocyanate (FITC) or tetramethylrhodamine (TRITC) were from Jackson ImmunoResearch Laboratories (West Grove, PA). Cell nuclei were visualized with 4',6-diamidino-2-phenylindole (DAPI) counterstaining.