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The amount of biotin was determined using the following equation: V biotin = 1, 000 × m PLGA M W biotin M W PLGA M biotin C biotin Open image in new window (2).
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The ratio of biotin to PLGA was determined using the recorded absorbance values.
The disruption force of specific biotin-streptavidin bonds was determined using DNA oligomers as force tags.
The concentration of biotin in the solution was determined using Fluorescence Biotin Quantification Kit Thermo Scientificc).
Since varying concentrations of avidin were used for coating the PCR tubes, optimized concentration of avidin needed was determined using biotin-HRP as shown in Figure 3.
Biotinylation grade was determined using a Biotin Quantification kit (Pierce) and protein concentration was determined using photometric analysis.
Band size was determined using pre-labeled biotin markers (Ambion).
The expression of VEGF and CD34 in the biopsy specimens was determined using the avidin-biotin complex method as described previously (Kaya et al, 2000).
The optimal incubation time was determined using the Streptavidin - Biotin-PE assay.
Thirdly, the optimal assay volume (0.5, 1, 2, 4, 8 and 16 µl) was determined using the Streptavidin - Biotin-PE assay.
The resulting 3' biotin surface densities were determined using the calibration of spotting concentrations with surface densities that was previously developed [ 13].
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