Ethancridine lactate added to the serum or plasma to give a last convergance of 5.2gm every liter. Pretty nearly 90% of the included sum was disposed of with the encouraged proteins, while the rest of which guarantees the complete precipitation of undesirable protein, uprooted by assimilation on charcoal. To sum things up, 1.2% ethancridine lactate arrangement was added to the serum or plasma and the mixture was mixed for 30-40 minutes at 200c and after that the mixture was kept undisturbed for 1-2 hours. After precisely tapping the supernatant the hasten was centrifuged at 5600 g for 30 minutes and the supernatant was pooled. To the supernatant charcoal at the rate of 6g/liter was added to ingest the broke up ethancridine lactate.
nilotica leaf extracts (0.3 mL) were mixed with 2.7 mL of methanol solution containing DPPH radicals (6×10-5 mol/L). The mixture has shaken vigorously and allowed to stand for 60 min in the dark. The reduction of the DPPH radical was determined by the absorbance at 517 nm (Barros et al. 2007). The radical-scavenging activity (RSA) was calculated as a percentage of DPPH discoloration, using the equation: %RSA = [(ADPPH - AS) /ADPPH] ×100, where, AS is the absorbance of the solution when the sample extract is added at a particular level and ADPPH is the absorbance of the DPPH solution.
After 10 minutes, the solution was deleaded by adding potassium oxalate crystals in excess and the volume was made up to a known amount with distilled water and filtered through whatman No. 1 filter paper. The filtrate was taken in a burette and titrated against boiling Fehling’s mixture (5 ml of Fehling’s solution A + Fehling’s solution 5 ml of B) till the blue colour faded. Then one ml of methylene blue indicator (1 per cent) was added and the titration was continued till the contents attained a brick red colour and titre value was noted. The percentage of reducing sugar was calculated according to the following
This test is based on removing of iron from chrome azurol S (CAS) by producing of sidrophore. Five microliters of each fresh culture was inoculated onto a plate, which was then inocubated at 28°C for 72 h. The presence of an orange halo (A change in a color of CAS from blue to orange) around a colony indicated a positive result (Schwyn and Neilands, 1987). 3.1.3 Indole-3-acetic acid production IAA production was assessed in tryptone broth medium ( tryptone 10.0 g, sodium chloride 5.0 g in 1000 ml distilled water and final pH 7.2). The medium was dispensed into tubes and autoclaved at121°C for 15 min. Bacterial isolates cultured for 24h were inoculated into tubes, which containing 5 ml tryptone broth and incubated at 37°C for 7days.
The other two concentrations were prepared from soaking sixfold aqueous methanol (75%) with different amounts of powder. At the end of extraction each extract was passed through the Whatman filter paper No. 1 (Whatman, UK). The methanol was removed from residue with rotatory evaporator (نام دستگاه، سال و .. ). Agar well diffusion method This method was used to determine the antibacterial activity of the extracts.
This organism was cultured under solid-state fermentation for 72 h using wheat bran as the substrate. After 72 h, crude enzyme was extracted from the culture medium. The fibrinolytic enzyme was purified from the crude sample by various steps: ammonium sulfate precipitation, dialysis, ion exchange chromatography, and casein-agarose affinity chromatography. All purification steps were performed at 4°C until otherwise stated. The crude enzyme was precipitated at 70% saturation of ammonium sulfate, and the protein was collected by centrifugation (10,000×g for 10 min).
1- Extraction method No. 1: Fifty grams of powdered aerial parts of portulaca oleracea were hydrolyzed by using reflux for 9 hr. with 300 ml of 2N hydrochloric acid then the extract cool at room temperature ,filter and wash the residue with 2N of ammonia solution. The residue dried overnight at 60ºC ,the final step involve the extraction of the residue with 250 ml of chloroform by using soxhlet ,the final extract cool at room temperature ,then filter and evaporate to dryness by using rotatory evaporator at 40ºC to yield (2.264 gm),as show in scheme (2-1). 220.127.116.11-Extraction method No.2: Fifty grams of powdered aerial parts of portulaca oleracea extracted by soxhlet with 500ml 0f 70% ethanol for 8 hr.
The residual biomass was separated by filtration and washed with distilled water. For alginate extraction, the acidified algal biomass was suspended in 3% Na2CO3 solution at different alkali: alga ratio (20, 40, and 60 mL/g). The different extraction temperatures ranged from 25 to 45º C, and lasted for 1 to 3 h. For each experimental run, sodium alginate was collected by filtration and precipitated with absolute ethanol (1:2 v/v). The mixture was maintained at 4º C overnight. The precipitate was collected by vacuum filtration and allowed to dry at room temperature.
In a separate beaker, 10-3 M of synthesized SB was dissolved in 10 ml DMF. The two solutions were mixed together under stirring and resulting yellow precipitate solution was transferred to a sonochemical bath. After 60 minutes of sonochemical treatment, the resulting CdS precipitate was collected, filtered, washed with double distilled water and absolute ethanol several times to remove the unreacted chemicals, and finally dried in an oven at 80oC for 5hours. Similar procedure was adopted to synthesize uncapped CdS
Plasmid DNA was precipitated from the supernatant by adding 0.7 volumes of isopropanol and centrifugation at 13,000rpm for 30min and washed with 70% ethanol. The pellet was air dried, resuspended in TE buffer (pH 8.0) with RNase 10μg/ml and allowed to dissolve completely. The isolated plasmid DNA was electrophoresed on 1 % agarose gel (containing Ethidium Bromide) at 75 volts and visualized under UV light in order to check its