Sentence examples for blocking buffer was added from inspiring English sources
Then, 100 μL of streptavidin-HRP (diluted at 1 5000 in PTM blocking buffer) was added into each well and incubated for 1 h at RT, 600 rpm to detect antigen-antibody binding.
Briefly, the provided blocking buffer was added to array membranes for 30 minutes.
Rabbit anti-LukF-PVL anti-serum (1∶20,000 in the blocking buffer) was added to each well.
A horseradish peroxidase-conjugated anti-mouse Ig (SouthernBiotech, Birmingham, AL, USA) diluted in blocking buffer was added and strips incubated for 1 h at 37°C.
diluted to 1/5000 in blocking buffer was added into each well and incubated for 3 hours at room temperature before detection with O-phenylenediamine (OPD, Sigma).
Two milliliters of 2% BSA blocking buffer was added to each tube, and tubes were rinsed by centrifugation at 350 g.
Primary antibodies rabbit IgG NRF2 (1 800, C-20, Santa Cruz), mouse IgG2b 8-OHdG (15A300, 15abcambcam) in blocking buffer were added to cells for 16 hours at 4 °C, when they were washed 3 times in PBS with 0.1% Triton X-100.
After blocking with 200 µl 1% BSA in PBST (phosphate buffered saline, pH 7.6, containing 0.05% Tween-20), 100 µl of prepared antibodies gradient diluted in blocking buffer were added and incubated at 37°C for 2 h.
Fluorescent secondary antibodies in blocking buffer were added and incubated for 1 hr at RT, followed by three washes in PBS for 5 min. Cells were visualized with a Olympus IX71 microscope with a CoolSnapHQ camera (Roper Scientific, Tucson/AZ, USA).
After washes, 50 µL of different concentrations of recombinant Hsp60 (ranging ranging from 20 µg/mL to 0.16 µg/mL in a 1∶2 serial dilution in blocking buffer were added to each well.
