Now heat to 80-85°C over 10 minutes and hold there for at least two minutes but not more than five. Cool in water to room temperature. Transfer the solution to a 100 mL beaker, keep some (~2-3 mL) in the Conical flask. Stir at medium speed on a magnetic stirrer. Now add 30% hydrochloric acid drop by drop till pH of the solution become 7 and filter it.
Fiver portions of about 3 mL of water was added in increments and cooled in an ice bath for about 10 minutes. After a new filter paper was add and the aspirin was placed in while the vacuum filtration was on and about 6 mL was poured in to get the last amount of
• The beaker was covered with the foil and stirred constantly on magnetic stirrer overnight (ideally 8 hrs) • After overnight extraction magnetic bead was removed and the extraction buffer was centrifuge... ... middle of paper ... ...ne (TMB) reagent • Stop solution 1. 2 N sulphuric acid • NUNC flat base ELISA wells PROCEDURE: • Prepare capture antibody solution of dilution 1:250 in coating buffer and add 100 µl in each well and incubate at 40C overnight. • Next morning aspirate and wash the wells 3 times with PBST • After washing add 200 µl blocking solution which is 10 % FBS in PBS in each well and incubate for 1 hour. • Aspirate and wash 3 times. • Add 100 µl standard and samples to the allotted wells.
Preparation of M. zapota seed extract: The seeds of M. zapota were collected freshly from the farms in Chennai, Tamil Nadu. The seeds were rinsed well with distilled water and air dried for two weeks before grinding them into a fine powder. The crude extract was prepared by dissolving 1gm of the powdered seeds in 10 ml of hexane, acetone and ethanol respectively under continuous shaking condition for 24 hours.The process is repeated thrice using fresh solvents. The solvent was removed by condensation and extracted residues were weighed in different solvents to yield 10mg/ml stock solutions. Screening of phytochemicals: The following tests were performed on the extracts to detect various phytoconstituents present in them with reference to
After incubation, the sample was centrifuged at 9.6K rpm for 10 min. Sample was then filtered with a .22μ filter before being loaded into HPLC for analysis. HPLC Method The HPLC method used was 20 μL of sample at 25°C through an RI detector for 15 minutes. The mobile phase was .0001M Sulfuric acid at .6ml/min with a column temperature of 60°C. Findings and Discussion: Preliminary Findings When samples first arrived, dilutions were made and tested on the HPLC.
After incubation, the mixture was then spinning at 13000 rpm for 30 minutes at 4 °C. The supernatant was discarded and wash with 70% cold ethanol. The pellet was allowed to air dry for 20-30 minutes and resuspend in water, nucleus free water or TE buffer.
2. Materials and methods Essential oil and mucilage extraction and purification For essential oil extraction, dry plant materials of rosemary distilled within 24 h in a steam distiller with an aqueous phase recycling system, using a plant material: water ratio of 2:1. The distillation time was about 2 h, and the oil obtained was separated from the aqueous solution and dried by treating with anhydrous Na2SO4. Each essential oil was transferred into a dark glass flask filled to the top and kept at a temperature of 4 °C until used (Meepagala et al., 2002). Also for mucilage extraction, Cactus stems were removed skin and cubed (1 cm3).
Rice straw and sugarcane bagasse were subjected to NaOH - microwave pretreatment and compared to untreated ones. Biomass (10%, w/v) was treated with NaOH solution for 30 min under boiling condition then subjected to microwave pretreatment under the optimized condition from the previous study. Biomass was washed several times in tap water followed by a final rinse in the distilled water. The lignocellulosic biomass was dried at 65 °C overnight. Finally, the pretreated biomass was cooled down at room temperature and stored in an airtight container until further use.
Protein quantification Five tubes labeled “Blank”, “C”, “LSS”, “HSS”, “HSP” were obtained. The blank contained 100 ul of distilled water. The others had 90 ul of distilled water and 10 ul of the cellular fractions obtained by centrifugation. #ml of BCA were added. The samples were placed in a water bath and incubated at 37 degrees Celsius for 30 min.
This was to see the difference between the initial temperature and the final temperature. First Test In a 250ml beaker place 100mls of water, measure the temperature of the water and record this initial temperature onto a table. Set the timer and add one teaspoon of Ammonium Nitrate to the water, stir this continuously until the Ammonium Nitrate has dissolved. After 1 minute measure the temperature and record it, do this for a further 2 minutes (3 minutes in total). Repeat this process for a total of 10 teaspoons.