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To verify the concentration, serial plating of the dilution (50 μl per plate and three plates per solution) was performed.
For the analysis of S100A1 serum concentration, serial blood samples were taken from the left carotid artery of anesthetized animals.
To estimate probe concentration, serial dilutions of labeled probes and a standard labeled RNA were spotted on nylon membranes (HybondN+; Amersham, USA), immunodetected using anti-DIG-Fab-AP conjugate diluted 1 5000, and visualized with NBT and BCIP according to instructions from Roche.
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A standard curve was made by preparing Blue Dextran 10 concentration serials and measuring with absorption at 620 nm.
The MEM was aspirated from the cells, and replaced with 200 μl of extract, fractions and compounds at differing concentrations (serial dilution prepared in growth medium).
All extracts were dissolved prior to use to the desired concentrations (serial dilutions from 1 mg/ml) in phosphate buffered saline (PBS) and stored at -20˚C.
Cells were treated the next day with tunicamycin (1 μg/ml) and different concentrations (serial dilution) of each compound for 7 hr.
The evaporated fraction was dissolved in absolute ethanol to 100 mg/ml, dissolved to desired concentrations (serial dilutions from 1 mg/ml) in PBS and aliquots stored at -20˚C until use.
A range of concentrations (serial dilutions 1 0.0625 m m) of Gd-DEG1SL, Gd-DEG3SL and Gd-DEG6SL were used to calculate relaxivity of each contrast agent as liposome formulations A – G.
One hundred μL of standard IPA concentration or serial dilutions of RG and AH and 100 μL diluted tracer were added to each well.
Observational management involves admission to a unit and the monitoring of vital signs, with strict bed rest, frequent monitoring of hemoglobin concentration and serial abdominal examinations [27].
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