First the solution of 1 mL of extract was added to deionizer water (10 mL), then Folin–Ciocalteu phenol reagents (1.0 mL) added to the mixture. The mixture was left for 5 minutes, and solution of 20% sodium carbonate (2.0 mL) was added to the mixture. This mixture diluted until 50 mL with deionizer water. And after that kept in total darkness room for 1 hour. The mixture absorbance was measured at 750 nm using a spectrophotometer.
Two ml of washed pack cells and 250 unit of FVIII were put into a dialysis bag (Sigma). Dialysis bag presoaked for 4 h in 50 ml PBS buffer containing 5 mM glucose. The bag was placed in a bea¬ker equipped with a magnetic stir bar. The beaker filled with 100 ml of hypoosmotic buffer containing 0.0451 mM KH2PO4/KOH, 0.1 mM MgCl2, 0.22 mM glucose, and 0.2 mM of adenosine triphosphate; pH 7.45. Dialysis was
Characterization of bacteriocins 6.1. Effect of temperature The effect of temperature on bacteriocin stability was determined by exposing 5 ml aliquots of CFS at 40C, Room temperature, 370C, 630C and 1000C for 30 minutes. Agar well diffusion method was performed to determine the residual activity in the heat-treated CFS17. 6.2. Effect of pH To check the pH stability of the bacteriocins, 5 ml aliquots of CFS was taken in sterile tubes and were adjusted to pH values of 2, 4, 7 and 9 busing 1N HCl or 1 N NaOH.
Three months later, the bone mineral density (BMD) of the femurs of all the rats were determined using GE Lunar Prodigy (GE Healthcare Bio Sciences, Pittsburgh, USA) to evaluate whether the model had been successfully established. Once the model is successful, the OVX rats were randomly divided into five groups of 14 animals: OVX group (double distilled water (0.2 mL/100 g, daily, administered orally)), positive group (70 mg/kg, weekly, administered orally) and OVX–HE groups (0.11, 0.33 and 0.99 g/kg, daily, administered orally) and treated for 12 weeks. The 0.11, 0.33 and 0.99 g/kg doses of HE were designated low (L-HE), medium (M-HE) and high (H-HE) groups,
Step-III: Synthesis of Cr(II) Complexes: The Schiff's base complexes were synthesized by mixing the Schiff's base (1.5 g) in ethanolic solution of Chromium chloride [CrCl2]. This reaction is refluxed in a waterbath for two hours and their volume were reduced to 70% of it’s original volume and residue was obtained. The coloured product obtained was filtered under suction, washed with ethanol. The product were recrystallized from ethanol. Their yields ranges from 50-55%, the product obtained were light green colour and melting point was 2100C.
Metal chelating activity Briefly, 2 mM FeCl2 was added to different concentrations of test sample and reaction was initiated by the addition of 5 mM ferrozine. The mixture was vigorously shaken and left to stand at room temperature for 10 min. Absorbance was measured at 562 nm after 10 min.8 % Inhibition = [(AB - AA)/AB] x 100, where AB, absorption of blank sample, AA, absorption of test sample. 2.6. Antibacterial
Heat the Agarose gel in a 65 °C water-bath to melt the agarose. After it melted, maintain its temperature at 55 °C until it is ready for use. 2. Transfer the spheroids from the 96-well plate to a 15 mL centrifugal tube using a 1000 μL pipette 3. centrifuge the tube for 5 min at 1000 rpm to form a pellet. 4.
Diabetes induced by injecting streptozocin bt introperitonial route of dose 60mg/kg . by dissolving it 0.9% normal saline solution. Diabetes was induced in two groups and remaing two as control group. Controlled group of rats was given only saline solution, while treated controlled animals was given with garlic extract at dose of 100mg/kg. This procedure was continued for three days.
This diluted solution will be used in the assay as duplicate samples. Then, 1.0mL of standard glycine solution containing (7.5mg/mL) was diluted to 100mL with water using a volumetric flask. This solution contains 1.0µmole/mL of glycine. 8 tubes were set up according to the following protocol and 2.0mL of ninhydrin reagent was then added to each of the 8 tubes and were placed in a boiling water bath for 20 minutes. After 20 minutes, the tubes from the bath was carefully removed, cooled in a beaker of cold water, then 8.0mL of 50% ethanol was added and mixed well.
The eggs were divided into 5 groups, each treatment was done in triplicates. Calamansi crude extract was reconstituted to three concentrations: (Treatment 1, 99μ of distilled water and 1μL of Calamansi crude extract) 1%, (Treatment 2, 95μ of distilled water and 5μL of Calamansi crude extract) 5%, and (Treatment 3, 90μ of distilled water and 10μL of Calamansi crude extract) 10%. Retinoic acid was used as a positive control, and ethanol as negative control. Approximately, 100μL of each treatment was placed in the filter paper. After the introduction of test solutions, the eggs were sealed with a tape and incubated for