Finding out the protein concentration in three different drinks to see if the food labels are telling the truth. The Bradford Assay experiment will be conducted to find out whether the protein concentration is true on the food label. We will be collecting the absorbances of each of these drinks and making a standard curve chart that will show which drink is high in protein. The FDA is the Food and Drug Administration and they are responsible for protecting the public health and making sure that
Chemistry Exploration Topic: determining the activation energy of a chemical reaction Research Question: What effect does temperature of the chemical reaction have on the activation energy ? ICT: Microsoft Word Autograph Microsoft Excel Introduction This experiment is designed to help in estimating the activation energy of the rate-limiting step in the acid catalyzed reaction of acetone with iodine. This is achieved by measuring the reaction rates at different reaction temperatures over
Enzyme assays are performed to serve two different purposes: (i) To identify a special enzyme by proving its presence or absence in a distinct specimen. (ii) To determine the amount of the enzyme in the sample by monitoring the disappearance of substrate or appearance of product. Enzymes speed up reaction rate by decreasing the activation energy required to start the reaction. Activation energy is the energy required to break certain bonds in the substrate so that other bonds can form. The formation
quantitative assay is a method used to measure the number of virus particles in an inoculated substance. There are various types of quantitative viral assays such as a plaque assay, infectious center assay, pock assay, and transformation assay. All of these assays contain different characteristics and are used to quantify different viral properties. In this lab, the quantitative assay that was constructed was a plaque assay. A plaque assay is a modification of a bacteriophage assay and is used to
Alamar Blue Assay detects cell viability through the compound resazurin in the Alamar Blue dye. Resazurin in its natural state is a blue non-fluorescing compound, but when reduced, resazurin fluoresces. This
The purpose of this experiment is to learn about the principles of protein assays as well as to learn how to utilize the Beer-Lambert Law by doing various calculations such as how to calculate absorbencies, concentrations, and extinction coefficients. According to the Beer Lambert Law, absorbance is proportional to path length and concentration. For this experiment we will be learning how to use a spectrophotometer which measures transmitted light intensity. Spectrophotometers measure wavelength
This purpose of this experiment was to determine if a new yeast transactivation assay was able to determine estrogenic activity in estrogenic compounds. Throughout this experiment, researchers compared the detection ability of the new transactivation assay with the conventional transactivation assay. The new transactivation assay plasmid replaced the lacZ gene with GFP (green fluorescent protein) gene, and by doing so simplifies the process of determining estrogenic activity. The advantages of using
2.5. Bovine Serum Albumin (BSA) binding Studies: BSA solution was prepared in 5 mM Tris-HCl/10 mM NaCl buffer solution to keep pH value constant (pH=7.4) and it is stored at 4oC, used within 2h. The BSA concentration was estimated spectrophotometrically (ε280=44,300 M-1cm-1) [35], 60 µM BSA was titrated with increasing concentrations of Ru (II) complex (0-50 µM). Absorbance changes at 280 nm were monitored by UV-spectrometry. 2.6. DNA Binding Studies: Binding studies of DNA were carried out at room
ACIDITY TEST INTRODUCTION: Acidity is the total amount of hydrogen ion present in the food sample with the expectation of those bound to alkaline ions. The hydrogen ion can be either attached to acids or in the form of free ions or anions. Titratable acidity is different than total acidity although at times both terms are used to mean the same thing total acidity is the total amount of organic acids in the food sample. This all acids (tartaric, oxalic acid, citric acid, sulfuric acid, lactic acid
Q1. explain briefly the difference between a) an agonist and antagonist –give 2 examples of each When the action of the drug is to activate or switch on a reaction, the drug is called an agonist. On the other hand, if the drug switches off the reaction, or inhibits or blocks the binding of other agonist components onto the receptor, it is called an antagonist. Hence, agonist and antagonist act in opposite direction, if agonist activate an action, antagonist inhibit an action(New Health Guide, 2014)
Acid phosphatase is an enzyme commonly found in nature in plants, animals, and protists. Acid phosphatase is found in human bone and prostate tissue. The optimal conditions of acid phosphatase were tested using Michaelis-Menten kinetics to determine the ideal substrate concentration, varied pHs were tested in order to determine the optimal pH, and various temperatures were tested in order to determine the optimal temperature. Using Michaelis-Menten kinetics, the Vmax was determined to be 312.5 nmol/min/mg
positive test for AO test will present a blue color, while a negative test will present no reaction. The flies in vial 1A will demonstrate AO activity while the flies in vial 1B will show no AO activity. So if the flies from vial 1A turn blue after the assay test, then they have the enzyme for AO activity. If the flies from vial 1B present no color change, then they do not have the enzyme for AO activity. In the second part of the lab, the pattern of inheritance of the aldox gene will be determined, whether
Acceptable criteria: % Assay should not vary by ±1%. 7.4.7. Specificity A solution of mixture of Amoxicillin trihydrate and its impurities was prepared by spiking of all three targetted impurities at a level of 0.01%. Acceptable criteria: Resolution of NLT 1.5 from primary peak
1.3.3 Methods of the measurement of DNase activity In 1950, the first method of the measurement of DNase I activity was described by Kunitz196. He isolated and precipitated DNase from fresh beef pancreas and isolated thymus nucleic acid. He found that the cleavage of DNA by crystalline DNase is accompanied by increase of absorption (at 260 nm) of UV light. This spectrophotometric method of measurement of the rate of the increase in the light absorption was then used for estimating of DNase activity
Isolation of Ecdysterone from Sesuvium portulacastrum As detailed in Figure1, Ecdysterone was isolated from Sesuvium portulacastrum using a sequential extraction process with Chloroform (CHCl3) followed by Methanol (MeOH), after which Alumina column chromatography of MeOH extract was performed. The different fractions were then eluted using varying proportions of CHCl3 and MeOH, resulting in MeOH: CHCl3 (20:80) bioactive fraction. Thin-layer chromatography (TLC) analysis of MeOH: CHCl3 (20:80) fraction
The antioxidant activity of the extracts was measured on the basis of the scavenging activity of the stable DPPH free radical (as cited in Dong et al., 2014). Evaluation of antioxidant activity of astaxanthin through DPPH assay was modified according to the procedures reported by Lewis (2012). 12 mls of 0.1 mM DPPH solution with methanol was prepared. A measurement of 0.005 g of DPPH was added to 12 mls of methanol which was measured with a graduated cylinder into a small
subcellular localisation of YAD in S. cerevisiae. The yeast cell was ruptured by homogenisation and fractionated by a process called centrifugation. Protein assay was carried out to calculate the concentration of protein prior to dilutions. ADH assay was carried out to oxidise the ethanol to acetaldehyde and two marker enzymes G6PDH and ALP assays were carried out to aid in the determination of the localisation on YAD. It is concluded that conclude that the ADH enzyme of yeast Saccharomyces cerevisiae
information Dot blot The Dot blot technique is a very simple method used to analyzing, detecting and identifying proteins. It is very similar in principle to the detection of samples in Southern blot, Northern blot, or Western blot. In the dot blot assay, the protein samples are not first separated by electrophoresis. Instead, the protein samples to be detected are spotted through circular templates directly onto the paper substrate or membrane. The Dot blot methodology offers significant savings
Testing and Counting with Trypan Blue Exclusion Method Anna Coretta R. Santos I. Introduction Cell viability assay plays a crucial role is in the cells of the cell culture. This assay can be used to identify the relationship of cell count from the cell’s behavior (Stoddart 2011). This test help researchers to determine the number of viable and non-viable cells in a cell culture. This assay helps in estimating the cell count of a cell culture and to determine if there is sufficient amount of viable
Since the 1950s, public support from animal testing has been in a steady decline and continues to drop every year. New developments are constantly being made throughout the world, so why are animals still being used as test subjects? Many cosmetic and hair companies do not test their products on animals; for example, Mac Cosmetics, Origins, Aveda, and so many other companies. However, there are still way too many companies that still do use animals to test their products. So many alternative ways