Sentence examples for before addition of a from inspiring English sources
A high specificity of the probe-receptor interactions could be demonstrated by blocking CCK2R binding positions with increasing concentrations of non-fluorescent, competitive minigastrin before addition of a fixed concentration of one of the optical probes.
The protein in PBS/2 mM EDTA (pH 7.0) was incubated for 30 min at 37°C with a 10-fold molar excess of TCEP (Bond-Braker, Pierce Biotechnology, Rockford, IL, USA) before addition of a 20-fold molar excess Alexa488-C5-maleimide Alexa488-C5-maleimide Alexa488-C5-maleimideture (RT).
The cells were allowed to settle for 10 min before addition of a coverslip.
Following stringent washing, 3′ end dephosphorylation was performed with T4 polynucleotide kinase (New England Biolabs) before addition of a pre-adenylated linker using RNA ligase (New England Biolabs).
Average fluorescence ratios were determined over 20-s periods, before addition of a test agent, during its perfusion, and after washout.
The solution was left to stir for 5 min before addition of a saturated solution of K2CO3 and extraction with CH2Cl2 (three times).
The nuclei were resuspended in 250 µl of NI buffer before addition of an equal volume of (10 mM HEPES [pH 7.9], 600 mM NaCl, 10% sucrose, 5 mM MgCl2, 0.1 mM EDTA, 0.5% NP-40, 1 mM DTT and protease inhibitor cocktail).
A volume of 50 μl was removed and frozen for analysis before addition of an equal volume of Surmodics Alk Phos Stabilzyme reagent.
Cells are allowed to settle for 1 h at 37°C before addition of an equal volume of 10% Matrigel in HuMEC media.
ADAMTS5-2 was incubated alone or with anti-ADAMTS-5 antibodies (total volume 5 μl) before addition of an equal volume gst-IGD flag substrate consisting of glutathione S-transferase (gst) conjugated with the IGD sequence of aggrecan core protein (Y G) and the C-terminal flag sequence (final concentration 17 μM) at 37°C for 30 min.
