TBP Monoclonal Antibody (1TBP18)

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	-Mouse (1:2000)

Protocol tips
-Mouse (1:2000)

Publication protocol

Nuclear fractions were prepared by incubation on ice in buffer A (25 mM HEPES, pH 7.6, 5 mM MgCl2, 25 mM KCl, 0.05 mM EDTA, 10% glycerol, 0.1% NP-40, 1 μM DTT, supplemented with protease inhibitors). After collection by centrifugation, pellets were resuspended in RIPA buffer (50 mM Tris, pH 8.0, 150 mM NaCl, 0.1% SDS, 0.5% sodium deoxycholate, 1% NP-40, 5 mM EDTA, 1 μM DTT, supplemented with protease inhibitors) and incubated on ice for 10 min. Whole cell lysates were prepared by incubation in RIPA buffer on ice for 10 min. Lysates were cleared by centrifugation and supernatants transferred to new tubes before measuring protein concentration by BCA assay. Equal protein amounts were added to loading buffer and samples then loaded onto 8–12% Bis-Tris gels. Following transfer, blots were incubated with primary and secondary antibodies before detection using Odyssey Imaging system (LI-COR Biosciences).

Antibodies
Primary antibodies used for western blotting are as follows: ARID1A (Santa Cruz Biotechnology, sc-32761), ARID1B (Abcam, ab57461), TBP (Thermo Scientific, MA1-21516), Brg1 (Santa Cruz Biotechnology, sc-17796), Brm (Bethyl Laboratories, A301-015A), BAF60A (Santa Cruz Biotechnology, sc-135843), BAF57 (Bethyl Laboratories, A300-810A), BAF53A (Novus Biologicals, NB100-61628), BAF180 (Bethyl Laboratories, A301-591A), MET (Cell Signaling Technology, #8198), V5 (Biorad, MCA1360). Antibodies used for ChIP: H3K27ac (Abcam, ab4729), H3K4me (Abcam, ab8895), H3K4me3 (Millipore, #05–745), H3K27me3 (Active Motif, #39155), FRA1 (Santa Cruz Biotechnology, sc-183x).

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