Membranes were blocked with 5% BSA in PBS for 1 h at room temperature and incubated with primary antibodies diluted 1:1,000 in blocking buffer overnight at 4 °C. Primary antibodies were: H3K9me1 (Upstate, 07-450), H3K9me3 (Millipore, 07-442) mouse anti-H3 (Abcam, ab10799), ...
Spots containing these purified proteins were then fixed in triplicate onto three respective rows for each membrane and dried at room temperature for ~20 min. The control membrane was blocked in a Petri dish with 5% (wt/vol) dry skimmed milk in TBS solution with 0.1% Tween (TBS-T), ...
Multisensory stimulation was performed as described previously6. In brief, mice were moved from the vivarium and held in a quiet room. Following 1 h of habituation to the room, individual mice were placed in separate chambers. The chamber was illuminated by a light-emitting diode programmed t...
Sections were permeabilized in 0.3% PBS-Triton for 15 min at room temperature, blocked in blocking buffer (5% donkey serum, 2.5% fish gelatin, 1% BSA, 0.3% Triton in PBS) for 15 min at room temperature. Primary antibodies in blocking buffer were incubated on slides overnight at 4 ...
Proteins were resolved by 4–12% gradient Bis-Tris SDS–PAGE (Invitrogen NP0323), transferred to PVDF membranes and blocked with 5% non-fat milk in TBST for 1 h at room temperature with shaking. The membranes were incubated with the following antibodies: anti-GAPDH (Abcam, ab59164, 1:2...
(1:250, cat# sc-166324, Santa Cruz), washed, and incubated for 45 min at room temp. in goat anti-mouse HRP (1:5000, cat# G21040, Life Technologies). Membranes were washed for 1 h at 4 °C, incubated in SuperSignal West Femto Chemiluminescent Substrate (Fisher Scientific), and ...
the inflow of gas from the room-temperature beamline is blocked, achieving an ultrahigh vacuum for long ion storage. For this measurement campaign, four hydrogen-like118Sn49+ions were loaded once. One of these was stored for three months, which allowed to precisely measure the magnetic moment ...
2 h later, cells were washed with DMEM to remove unbound mAbs and then treated with 0.1 M citrate pH 5.0 for 5 min at room temperature to induce M-mediated cell-cell fusion. After this, the acidic medium was replaced with fresh DMEM supplemented with 4% FBS and 10 mM HEPES...
Endogenous peroxidase was blocked using 3.0% hydrogen peroxide for 10 min. Nonspecific background blocking was performed using Background Punisher (Biocare Medical, Pacheco, CA). The sections were incubated for 1 h at room temperature in Rabbit polyclonal anti-NELF-B rabbit polyclonal antibody ...
7). At room temperature, a voltage applied to the interdigitated electrodes initiates the reaction 2H2O (liquid) → O2 (gas) + H2 (gas). The resulting gas pressure deforms the flexible membrane, thereby driving the flow of liquid from the μ-reservoir through the passive flapper...