Anti rbpj antibody

Satellite cells were isolated from adult Tg:Pax7-nGFP mice using in situ fixation to preserve Notch signalling from dissociation-induced downregulation39 . Cells were fixed as above in (2mM Di(N-succinimidyl) glutarate for 45min, followed by 10min with 1% formaldehyde at RT). Cell lysis and chromatin isolation were performed using Auto-TrueMicrochip kit (Diagenode, C01010140). Chromatin was sheared as above with 10 cycles of 30s on/off sonication using a Bioruptor Pico. 2x10⁵ cells were used per ChIP and 2x10³ per Input and IPs were performed using 2μl of anti-Rbpj antibody (Cell Signalling, 5313) or 0.5μl of rabbit control IgG antibody following the manufacturer guidelines. Immunoprecipitated chromatin preparations and input were purified using the Auto IPure kit v2 (Diagenode). RT-qPCR was performed using FastStart Universal SYBR Green Master mix (Roche, 04913914001) and analysis was performed using the 2^-∆∆CT method38 (link) normalised to the Neg16 region. Primers used for ChIP-qPCR are listed in Supplementary Table 1.