After cooling, the solution was removed.
After incubation, the solution was removed and the cells were washed with PBS for at least three times.
Then, the solution was removed from the heat, and the stirring process continued for another 10 15 min.
At the end of the 4 h incubation, the solution was removed and the wells were washed three times with BSA/PBS.
After a predetermined time, the solid and liquid from one flask were separated; after the next period, the solution was removed from the second flask.
An aliquot of the solution was removed and diluted with Griess reagent (1 % sulfanilamide in 5%H3PO4and0.1%0.1 % naphthylethylenediamine dihydrochloride).
An amount (3 ml) of the solution was removed from the beaker every 10 min to test the absorbance properties at 464 nm after filtrating the solid spheres.
Following an overnight incubation at 4°C, the solution was removed and 1% bovine serum albumin (BSA) in phosphate-buffered saline (BSA/PBS,150 μL) was added to each well for 1 h at ambient temperature.
The solution was removed, and the cells were washed with sterile PBS and treated with varying concentrations (ranging from 10too 100 µM) of Pep-A and B for 24 h.
The solution was removed and serial dilutions (1,000-17 ng) of the panitumumab F ab' 2 (50 μL) were added to the wells in triplicate; one set of wells received only BSA/PBS.
Untransformed MTT in the solution was removed by aspiration.
