The ligand binding assay was a variant of the ELISA in which dilutions of ligand (rhFGF-2) were allowed to bind to a ligand binding protein such as rhPln.D1 that had been coated onto the wells in PBS-N3.
The biotinylated peptides (20 pmol in 50 µl ddH2O) were allowed to bind the streptavidin-coated high-capacity binding plates (Pierce #15,503, Rockford, IL) overnight at 4°C or 2 hr at room temperature.
32P-labeled (1 pmol, 20,000 cpms) IDX-rasISS1 RNA was allowed to bind to 640 ng recombinant hnRNP H in binding buffer (1 mM ATP, 1.7 mM MgCl2 and 0.1 µM heparin in 1×Roeder D buffer).
After addition of a binding buffer and the magnetic particles (Siemens Healthcare Diagnostics, Cologne, Germany) nucleic acids were allowed to bind to the particles for 15 min at room temperature.
Then the mixtures were allowed to bind to glutathione-Sepharose.
Antiserum samples after different consecutive immunizations were allowed to bind to the immobilized antigen (BSA and ROS-BSA).
Antiserum samples after different consecutive immunizations were allowed to bind to the immobilized antigen (BSA and LND-BSA).
Then an enzyme labeled detector anti-cTnI antibody is allowed to bind with the immobilized cTnI.
Note that the players are allowed to bind an agreement only on the space of feasible outcomes.
The supernatant was added to a Ni2+-NTA resin column (1 ml bed volume) equilibrated with lysis buffer and was allowed to bind slowly.
