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Concentrations of each soluble adhesion molecule were quantified using commercially available ELISA kits [R&D Systems plc, Abingdon, UK].
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In the present study fluorescence patterns from fewer single molecules were quantified due to the more extensive analysis needed for each pattern.
Small molecules were quantified using previously determined calibration curves.
Small molecules were quantified using a previously determined calibration curve.
All the molecules were quantified using enzyme-linked immunosorbent assay (ELISA).
The level of released total aggrecan molecules were quantified using the G1/G2 assay.
After washing, bound gliadin molecules were quantified with an ELISA assay using the commercial human anti-gliadin IgG antibodies.
The numbers of biotinylated TGF-β1-bound TβRII molecules were quantified using biotinylated human TGF-β1 (R&D Systems), according to the manufacturer's instructions.
Snap-frozen postmortem liver and skeletal muscle biopsies were homogenized and protein levels of signaling molecules were quantified with immunoprecipitation (IRS 1+ PI3K and SHC + Grb2), western blotting (phosphorylated Akt) and ELISA (phosphorylated p42/44MAPK).
Samples were taken from different joint areas after 6, 12, 24 and 48 weeks, and gene expression levels of common cartilage molecules were quantified in relation to the histological grading (modified Mankin score) of adjacent tissue.
No further increase in the number of molecules were observed once all molecules were quantified, and the ratio of observed to expected molecules stabilized around 1. Increasing the annealing and elongation time also reduced the impact of SNPs (see Additional file 1).
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