The feed sample and the resultant fractions, produced by each separation technique, were analyzed with ICP-MS to determine the zirconium and REE content.
The resultant fractions were then analyzed for molecular-structure information via matrix-assisted, laser desorption/ionization (MALDI) mass spectrometry and by high performance liquid chromatography (HPLC) with UV vis detection.
The resultant fractions of the gradient were processed to SDS-PAGE and Western blotting for detection of IC74.
The resultant fractions were analyzed using an Orbitrap mass spectrometer.
The resultant fractions were assayed for neurite outgrowth in PC12 cells based on microscopic assessment.
SDS-PAGE analysis of the resultant fractions revealed that the higher-order multimers could be separated from the homodimeric fractions with a relatively high degree of purity.
Whole cell extracts were separated on 10%40%% glycerol gradients, and the resultant fractions analyzed by gel electrophoresis followed by western/northern blotting or, in the case of labeled RNAs, phosphorimager analysis.
The resultant soluble fraction was removed and designated the soluble cytosolic fraction (S).
The resultant P2 fraction was resuspended in 250 µl APT-1 reaction buffer containing PIM/PMSF and subjected to acyl-protein thioesterase treatment by incubation with recombinant APT-1 as previously described [13], [14], [25] with some modifications.
Briefly, after low-speed centrifugation, the supernatant was collected and further centrifuged at 100,000×g for 30 min at 4°C and the resultant membrane fraction was resuspended in 10 mM Tris pH 7.5, 0.1 mM EDTA and stored at −70°C.
