3.2.1 Preparation of Plant Extract
Extraction of Nypa Fruticans fruit is carried out using soxhlet extraction method. The aim of this method is to extract solid samples by manipulating the difference in solubility to separate desired compound while leaving the undesired compound. For example, in a salt and sand mixture the solubility of each of them is different. When water is added as the solvent and the solution is stirred, salt will dissolve and sand will stay as it is insoluble. The same principle applies in soxhlet extraction method. It is carried out using 45g of samples and 500 ml of solvent. In this study, two types of solvent are used which re methanol and n-hexane. Like dissolve like theory is reflected in the process. Methanol…show more content… A first anibody known as the capture antibody will be added to the microwell plate and it adsorbed to the well surface. The purpose of this antibody is to bind with the target antigen. Then samples are added to the well and binding process between the first antibody with the target antigen occurs. The well is left for 1 to 2 hours for the binding to take place properly. The microwell plate is washed and a second antibody or called as detection antibody, is added to the same well to recognize the bound complex. This detection antibody is conjugated with an enzyme, usually horseradish peroxidase. It catalyzes the hydrolysis of chromogenic substrate which is added later. The chromogenic substrate undergoes colour change and it is determined using spectrophotometric plate reader at the specified wavelength. Before the absorbance for each microtitre plate well is measured a stop solution is added to the mixture to stop the reaction. ELISA kit for adiponectin from ABCAM will be used to carry out this procedure and the instruction included in it will be followed. Adiponectin is the target antigen so an antibody which is specific to adiponectin will bind to it and the level or amount can be measured