C notch1 4147

Whole cell lysates (15 μg/lane) were resolved in 4% to 20% SDS-PAGE from Bio-Rad (Hercules, CA). Blots were then probed with the indicated antibodies. Primary antibodies against c-Notch1 (4147, 1:1000), cytokeratin 18 (4548, 1:1000) and Oct4 (2750, 1:1000) were from Cell Signaling Technology (Beverly, MA); β-actin (A1978, 1:2000) was from Sigma-Aldrich (St. Louis, MO). Secondary antibodies were from Cell Signaling Technology. Western blot images are quantified by using GeneGnome XRQ chemiluminescence imaging system and analyzed by GeneTools analysis software from Syngene (MD, USA).

The procedures have been described previously (25 (link)). Whole-cell lysates (15 μg/lane) were resolved in 4% to 20% SDS-PAGE from Bio-Rad (Hercules, CA). Blots were then probed with the indicated antibodies. Primary antibodies against CD44 (sc-7297, 1:200), which recognizes both CD44s (standard form) and CD44v (variant form), NFκB (sc-372, 1:200) and cyclin D1 (sc-718, 1:200) were from Santa Cruz Biotechnology (Santa Cruz, CA); CD49f (3750, 1:1000), Notch1 (4380, 1:1000), c-Notch1 (4147, 1:1000), and pNFκB (3031, 1:1000) were from Cell Signaling Technology (Beverly, MA); CD24 (555426, 1:200) was from BD Biosciences (San Jose, CA), proliferating cell nuclear antigen (PCNA) (M0879, 1:400) was from Dako (Carpinteria, CA), and β-actin (A1978, 1:2000) was from Sigma-Aldrich (St. Louis, MO). Secondary antibodies were from Santa Cruz Biotechnology.