The experiment calls for the extraction and purification of a protein. The protein chosen for the experiment was an amylase, specifically beta-amylase. Amylase protein can be typically found in plants and animals. The role of this protein is to hydrolyze organic materials, such as starch and dextrin, in order to form glucose, maltose, and limit dextrin. One distinction between beta amylase and alpha amylase is that beta amylase is not found in animal. It is found primarily in plants, bacteria, and fungi. In these types of organisms, beta amylase plays a huge role in its survival. It contributes to the hydrolysis of glycosidic bonds. Also, in plants like sweet potatoes (source of the experiment), the protein performs the hydrolysis of starch
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Like the sodium phosphate buffer, the solid was dissolved in 300 milliliters of distilled in a 500-mL beaker using a stirrer and a magnetic stirring bar. The pH of the solution was then checked using the pH meter and it was brought down to the desired pH of 4.8 using a couple of drops of glacial acetic acid. Then it was poured into a 500-mL graduated cylinder and brought up to a volume of 1000 milliliters using distilled water. The buffer was poured into a labeled glass bottle and stored in the fridge. Another sodium acetate buffer was made using the same …show more content…
Each fraction was prepared in an eppendorf tube and the solutions were added using a 100uL-1000uL micropipette and micropipette tips. The first was the blank solution that only contained 800 microliters of distilled water and 200 microliters of the BioRad protein assay reagent. The second fraction was made with 200 microliters of distilled water, 600 microliters of the BioRad protein assay reagent, and 200 microliters of Bovine serum albumin standard solution (BSA). The following fractions had the amount of water used decrease by 200 microliters and the amount of BSA solution used increase by 200 microliters. The amount of BioRad reagent used in each fraction was kept constant. Each fraction had a total volume of 1000 microliters or 1
Like the sodium phosphate buffer, the solid was dissolved in 300 milliliters of distilled in a 500-mL beaker using a stirrer and a magnetic stirring bar. The pH of the solution was then checked using the pH meter and it was brought down to the desired pH of 4.8 using a couple of drops of glacial acetic acid. Then it was poured into a 500-mL graduated cylinder and brought up to a volume of 1000 milliliters using distilled water. The buffer was poured into a labeled glass bottle and stored in the fridge. Another sodium acetate buffer was made using the same …show more content…
Each fraction was prepared in an eppendorf tube and the solutions were added using a 100uL-1000uL micropipette and micropipette tips. The first was the blank solution that only contained 800 microliters of distilled water and 200 microliters of the BioRad protein assay reagent. The second fraction was made with 200 microliters of distilled water, 600 microliters of the BioRad protein assay reagent, and 200 microliters of Bovine serum albumin standard solution (BSA). The following fractions had the amount of water used decrease by 200 microliters and the amount of BSA solution used increase by 200 microliters. The amount of BioRad reagent used in each fraction was kept constant. Each fraction had a total volume of 1000 microliters or 1