Sentence examples for blocking buffer was applied from inspiring English sources
For CBHI's Western blot, first 4 μL of monoclonal anti-CBHI antibody in 5 mL SuperBlock T20 phosphate-buffered saline (PBS; pH 7.4) blocking buffer was applied, followed by 4 μL secondary anti-mouse antibody in 5 mL SuperBlock T20 PBS blocking buffer.
After washing, 100 µL of each serum previously treated with concanavalin and diluted 1∶50 in blocking buffer was applied to each well.
One µg/mL each of anti-TfR1 (Invitrogen) or anti-actin (Sigma) antibody, diluted in blocking buffer, was applied to the blot for 1 hour at room temperature.
Subsequently, 5 % normal goat serum blocking buffer was applied.
In brief, primary anti-Ntrk2 antibody (Abcam, ab33655) diluted 1 200 in the blocking buffer was applied to 10 um thick mouse lung sections and incubated for 30 min.
After removing the blocking buffer from the plate, 20 μL of serum of 200 μg/mL total protein in blocking buffer was applied to each well and the plate was incubated at room temperature for 2 h with gentle shaking.
After extensive wash the secondary antibodies (in blocking buffer) were applied for 1.5 hours at RT in darkness.
Either rat anti-mouse CD31 or isotypic antibodies 1∶100 in 10× diluted blocking buffer were applied for 1.5 h and then sections were washed with PBS.
Primary antibodies in blocking buffer were applied to the cells for 1 hr at RT, followed by three 5 min washes in PBS.
Phalloidin-Alexa Fluor 568 (Invitrogen A12380, diluted 1 50 in blocking buffer) and/or BrdU-FITC (Becton-Dickinson 347583, diluted 1 4 in blocking buffer) were applied for 30 min before final washes and mounting in Vector-shield containing 0.5 mg/ml DAPI (Sigma D9542).
After blocking with 5% normal goat serum for 30 min, primary DAB2IP polyclonal antibody (1 50, gift from Prof. J. T. Hsieh) as well as Tp53, Ki67, E-cadherin, and vimentin polyclonal antibody (all 1 100; Santa Cruz Biotechnology, Santa Cruz, CA, USA) in blocking buffer were applied and the slides were incubated overnight at 4°C.
