The pH value shows a linear change with respect to the addition of NaOH both before and after the reaction, which is due to the buffer behavior of the sodium citrate and the low alkali dosage.
The performance of NCBs was evaluated by comparing their bulk enhancement ratios, which are defined as ER = (Iactivated – Ibuffer)/(Iinactivated – Ibuffer), where Iactivated, Ibuffer, and Iinactivated are the integrated fluorescence intensities of the activated NCB (i.e., the duplex), the sodium phosphate buffer, and the inactivated NCB (i.e., NC probe only), respectively.
The sodium acetate buffer had a final concentration of 50 mM sodium acetate with 100 mM sodium chloride and 10 mM magnesium chloride and the sodium phosphate buffer had a final concentration of 50 mM sodium phosphate with 100 mM sodium chloride and 0.1 mM EDTA.
At an age of eight weeks, half of the animals were injected intraperitoneally with streptozotocin 60 mg/kg dissolved in sodium citrate buffer, and the other half with buffer alone.
The reaction was stopped by replacing the substrate with sodium acetate buffer and the results were examined with a microscope.
The fixative was subsequently decanted and replaced with 0.1 M sodium cacodylate buffer and the sample re-suspended and left for 2 hours.
For 4,6-αGT-W activity, the optimal pH and temperature were determined over the pH range of 3.5 6.5 (in 25 mM sodium acetate buffer) and the temperature range of 20 60°C, using 100 mM maltose as substrate by following the release of glucose in time using the GOPOP kit (Megazyme) (Kaper et al. 2007).
For the glycerol assay, aliquots of each sample (300 μl) were mixed with 300 μl sodium phosphate buffer, and the samples were measured with a Bruker AVANCE III NMR spectrometer (Bruker Biospin, Rheinstetten, Germany) operating at 500.36 MHz using a Bruker 1D CPMG pulse sequence (Bruker Biospin) with water peak suppression.
Three parameters were selected for this study: the buffer pH, the buffer concentration and sodium dodecyl sulphate (SDS) concentrations.
These pairs included creatinine and blood urea nitrogen (BUN) (both related to renal function), bicarbonate and lactate (related physiological because the former acts as a buffer for the latter), sodium and chloride (both constituents of a common resuscitation fluid), and white blood cell count (WBC) and platelet count (hypothesized to contain less redundancy).
