Protocol detail

Chromatin Immunoprecipitation Protocol

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Chromatin was prepared and immunoprecipitation was undertaken according to protocols described previously (Sofueva et al. 2013 (link)). Sonication was performed in 0.7% SDS using a Diagenode Bioruptor (maximum power 30 sec on and 30 sec off for 45 min). Pull-down was undertaken using Dynabead protein G sepharose beads (Thermo Scientific) conjugated with 10 µL of ChIP-grade antibody (anti-FoxG1 [Abcam, ab18259] and anti-V5 [Abcam, ab15828]) diluted in 250 µL of buffer.

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Bulstrode H., Johnstone E., Marques-Torrejon M.A., Ferguson K.M., Bressan R.B., Blin C., Grant V., Gogolok S., Gangoso E., Gagrica S., Ender C., Fotaki V., Sproul D., Bertone P, & Pollard S.M. (2017). Elevated FOXG1 and SOX2 in glioblastoma enforces neural stem cell identity through transcriptional control of cell cycle and epigenetic regulators. Genes & Development, 31(8), 757-773.

Publication 2017

Diagenode Bioruptor ab18259 ab15828

Antibody Buffer Chip Chromatin Immunoprecipitation Protein g Pull Sepharose

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Variable analysis

independent variables

Sonication conditions (0.7% SDS, Diagenode Bioruptor, maximum power 30 sec on and 30 sec off for 45 min)

dependent variables

Chromatin immunoprecipitation (ChIP) with anti-FoxG1 and anti-V5 antibodies

control variables

Dynabead protein G sepharose beads
ChIP-grade antibody (anti-FoxG1 [Abcam, ab18259] and anti-V5 [Abcam, ab15828]) diluted in 250 µL of buffer

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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