Discussions Bradford reagent is used to determine the concentration of protein in the experiment, it is a red brown acidic solution in the presence of protein. Bovine serum albumin (BSA) used for various biochemical applications include ELISA which is Enzyme-Linked Immunosorbent Assay, high content of screening assays, western blotting and immunohistochemistry. Furthermore, Bovine Serum Albumin can also be used to determine the quantity of other proteins which is by comparing an unknown quantity of protein to known amount of BSA. In this experiment, Bovine serum albumin (BSA) solution is used as protein concentration standard as the stability of bovine serum albumin will increase signal in the assays and lack of effect in biochemical reactions. Both of these solution are used in part 1 and part 2. A graph of absorbance against volume of BSA protein concentration is plotted in part 1. A graph is shown approximately whereby the absorbance increases when the volume of BSA concentration increases. Since the …show more content…
Thus, the larger the molar absorptivity, the more probable the electronic transition. In the UV spectroscopy, the concentration of the sample solution is measured in molL-1 and the length of the light path in cm. Hence, given that absorbance is unitless and the unit of molar absorptivity are L mol-1 cm. Since the units of molar absorptivity is always the above, it is customarily reported without unit. The derivative of the Beer-Lambert law is shown below: Step 1: Assumption one relates the absorbance to concentration and can be expressed as A∝ c (1) Step 2: The absorbance (A) is defined via the incident intensity Io and transmitted intensity I
Data from Table 1. confirms the theory that as the concentration of glucose increases so will the absorbance of the solution when examined with the glucose oxidase/horseradish peroxidase assay. Glucose within the context of this assay is determined by the amount of ferricyanide, determined by absornace, which is produced in a one to one ratio.1 Furthermore when examining the glucose standards, a linear calibration curve was able to be produced (shown as Figure 1). Noted the R2 value of the y = 1.808x - 0.0125 trend line is 0.9958, which is statistically considered linear. From this calibration curve the absorbance values of unknowns samples can be compared, and the correlated glucose concentration can then be approximated.
Data table 1 Well plate Contents Glucose concentration A 3 drops 5% sucrose + 3 drops distilled water Negative B 3 drops milk+3 drops distilled water Negative C 3 drops 5% sucrose +3 drops lactase Negative D 3 drops milk +3 drops lactase 15+ E 3 drops 20% glucose +3 drops distilled water 110 ++ Questions B. In this exercise, five reactions were performed. Of those reactions, two were negative controls and one was a positive control.
Absorbance was defined as: log I_o/I where I_o is incident light and I is the transmitted light. Fluorescence emission spectrum is different from fluorescence excitation spectrum because it records different wavelengths of chemical s...
The analyzed yellow#5 wavelength was determined to 395nm because the actual wavelength 427nm was restricted in the Micro lab. The R2 value of the graph is 0.9827, and the level of data accuracy it indicated extremely weak data correlation. The first one dilution data points excluded from the standard curve because the point is not in the linear curve. The first concentration and absorbance value are the highest point in the graph that cannot connect as linear with another data point. After removing the first data point, the standard curve is clear and make
The affects of pH, temperature, and salt concentration on the enzyme lactase were all expected to have an effect on enzymatic activity, compared to an untreated 25oC control. The reactions incubated at 37oC were hypothesized to increase the enzymatic activity, because it is normal human body temperature. This hypothesis was supported by the results. The reaction incubated to 60oC was expected to decrease the enzymatic activity, because it is much higher than normal body temperature, however this hypothesis was not supported. When incubated to 0oC, the reaction rate was hypothesized to decrease, and according to the results the hypothesis was supported. Both in low and high pH, the reaction rate was hypothesized to decrease, which was also supported by the results. Lastly, the reaction rate was hypothesized to decrease in a higher salt concentration, which was also supported by the results.
One of the most primitive actions known is the consumption of lactose, (milk), from the mother after birth. Mammals have an innate predisposition towards this consumption, as it is their main source of energy. Most mammals lose the ability to digest lactose shortly after their birth. The ability to digest lactose is determined by the presence of an enzyme called lactase, which is found in the lining of the small intestine. An enzyme is a small molecule or group of molecules that act as a catalyst (catalyst being defined as a molecule that binds to the original reactant and lowers the amount of energy needed to break apart the original molecule to obtain energy) in breaking apart the lactose molecule. In mammals, the lactase enzyme is present
Thyroid and metabolism hormones play a large role in the daily lives of all living species. Thyroid hormones regulate the metabolism and the metabolism is responsible for maintaining a specific range for the biochemical reactions that occur in the body (Martini 2014). The most important hormone for metabolic maintenance is thyroxine (T4). This hormone also plays a large role in body heat regulation. It is produced by the pituitary gland and secreted by the thyroid gland. The thyroid releasing hormone (TRH) must trigger the thyroid stimulating hormones (TSH) to release thyroid hormones to the thyroid gland. These hormones are under control of the hypothalamus, or main neural control center. Propylthiouracil (PTU) is a medication used to treat
...eases, including temperature. It is determined from the data that the reaction is more likely to have a step wise mechanism than a concerted due to the small – ΔS and a relatively large value of ΔH from the tables. Due to some errors, it is best to perform another experiment for future protocols. In addition with the variance the 35°C where at one point the absorbance levels off and then increases. In comparison to the rate constant against temperatures, at 25°C it is higher than 35 and 45. More test is required to ensure proper determination of the rate constant at those temperatures.
Mader, S. S. (2010). Metabolism: Energy and Enzymes. In K. G. Lyle-Ippolito, & A. T. Storfer (Ed.), Inquiry into life (13th ed., pp. 105-107). Princeton, N.J: McGraw Hill.
The least absorbed was the 4 Celsius with only 0.040 absorbed. The most absorbed was -20 Celsius with 0.219 absorbed.
The independent variable for this experiment is the enzyme concentration, and the range chosen is from 1% to 5% with the measurements of 1, 2, 4, and 5%. The dependant variable to be measured is the absorbance of the absorbance of the solution within a colorimeter, Equipments: Iodine solution: used to test for present of starch - Amylase solution - 1% starch solution - 1 pipette - 3 syringes - 8 test tubes – Stop clock - Water bath at 37oc - Distilled water- colorimeter Method: = == ==
...le that has an intensity of 10-3 W/m2 can be determined2: db = 10 log 10 ( 10-3/10-12 ) = 90-dB.
...eadings. The absorbance readings for test tube 5, were always further away from the expected values than test tube 1. This is because the NaOH was not added to each tube at a time, but in sequential order with the test tube numbers. This allowed the reaction in test tube 5 to proceed longer than in test tube 1, allowing more product to be produced, giving a higher absorbance reading than expected. In fact, this trend was shown in all the test tubes. In increasing order of test tube numbers, every absorbance was more off than expected.
The system involved in this lab was L-dopa as a substrate, enzyme was Tyrosinase, and the product was Dopachrome. Tyrosinase is commonly known as polyphenol oxidase, an enzyme that present in plant and animal cell (#1 Boyer). In plant cell, the biological function if Tyrosinase is unknown, but its presence is readily apparent. Tyrosinase is also involved in the browning of fruits, tubers, and fungi that have been damaged. In mammalian cell, Tyrosinase is involved in melanin synthesis, which gives skin its color. It will act on the substrate L-dihydroxyphenylalanine (L-Dopa) and convert to Dopachrome, which is the product that has color, and it can measure at 475nm using the Spectrophotometer. This work based on the Beer-Lambert’s Law (A=εlc), A stands for Absorbance, ε is extinction coefficient or the molar absorptivity (M-1 cm-1), and l is the path length (distance) that light passes through the sample (cm), c is a concentration of solution (M) (#3 Ninfa, Ballou, Benore). Beer- Lambert Law predicts a linear relationship between absorbance and the concentration of a chemical species being analyzed. It states that the absorbance (A) of a sample solution is directly proportional to the concentration (c) of the absorbing colored
= Before conducting the experiment I would conduct a simple test for the protein by placing a sample of the albumen into a test tube and add biurett reagent. This contains copper (II) sulphate and sodium hydroxide.