Streptavidin alexa fluor790

Protein samples were separated by SDS–PAGE using a 4–20% gradient gel (Bio‐Rad) and then transferred onto a 0.45 μm nitrocellulose membrane (Pall Corporation) using a Trans‐Blot Turbo transfer system (Bio‐Rad). Membranes were blocked in 2% wt/vol Bovine Serum Albumin (BSA) in phosphate‐buffered saline (PBS) solution for 30 min at room temperature (RT), incubated for 1 h at RT with rabbit anti‐Histone H3 (ab1791, 1:5,000; Abcam) diluted in a 2% wt/vol BSA/PBS solution containing 0.01% NaN3. After washing, membranes were then probed with secondary goat anti‐rabbit‐IRDye680RD antibody (ab216777; Abcam) and streptavidin‐Alexa Fluor790 (S11378; Invitrogen), both diluted 1:10,000 in 2% wt/vol BSA/PBS solution for 1 h at RT. Blots were washed and imaged on the LI‐COR Odyssey Infrared Scanner. Images were quantified using GelAnalyzer 19.1 (Lazar & Lazar, n.d. ).

The lysate was blotted to a nitrocellulose membrane (pall) using the RoToR into a 1,536 dot array using the 384 long pads (Singer Instruments) calibrated to move 200 pl of liquid in three technical replicates. The membrane was dried overnight at RT and blocked for 30 min with 2% (W/V) BSA in PBS (Sigma), incubated while shaking ON at 4°C with rabbit anti‐Histone H3 (Abcam) antibody used as a control, washed, developed using goat anti‐rabbit‐IRDye680RD antibody (ab216777; Abcam), and streptavidin‐Alexa Fluor790 (S11378; Invitrogen), on an odyssey LI‐COR imaging system.