Immunoblotting

GD Grzegorz Dobrynin
TM Tom E. McAllister
KL Katarzyna B. Leszczynska
SR Shaliny Ramachandran
AK Adam J. Krieg
AK Akane Kawamura
EH Ester M. Hammond
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Cells were lysed in UTB (9 M urea, 75 mM Tris-HCl pH 7.5, 0.15 M β-mercaptoethanol) and briefly sonicated. Primary antibodies used: HIF-1a (BD Transduction Labs., 610959), KDM4A (Abcam, ab24545), CAIX (BioScience, AB1001), Glut1 (Abcam, ab14683), Actin (Santa Cruz, sc-69879), H3K9me3 (Millipore, 07-422), H3K36me3 (Abcam, ab9050), H3 (Cell Signaling, 36385), NFκB p52 (Millipore, 05-361), Sp1 (Millipore, 07-645), E2F-1 (Cell Signaling, 3742S), HIF-2a (Novus Biologicals, NB100-122). Secondary antibodies were IRDye® 680RD Goat anti-Mouse IgG (H+L), IRDye® 680RD Goat anti-Rabbit IgG (H+L), IRDye® 800CW Donkey anti-Mouse IgG (H+L) and IRDye® 800CW Donkey anti-Rabbit IgG (H+L) from LI-COR Biosciences. Odyssey IR imaging technology (LI-COR Biosciences) was used for imaging.

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