...4PY013 MOLECULAR BASIS OF LIFE PROTEIN ASSAY BY BIURET AND FOLIN METHODS Student Number and Name:1316740 Katerina Koupa | Name of Lab Instructor: Dr. J. Walton | Date of the experiment:06/02/2014 | Date of Report Submission:20/02/2014 | (Fig.1) Biuret Reagent Structure ABSTRACT: This report describes the use of the Biuret and Folin methods of protein assay, to verify the protein concentrations of two unknown solutions. To calculate the protein concentrations of the unknowns, nineteen samples were passed through a spectrophotometer which twelve of them had a known protein concentration and there absorbance levels were found. Then a calibration graph was plot that determines the concentration levels of the unknown samples. INTRODUCTION: One of the most typical procedures applied by lab scientist is the protein quantification. Finding protein concentrations in solutions is important in many ways. First of all detecting the levels of a protein in body fluids can result in identifying various diseases. Also protein verification is required for characterization and purification of enzymes. Because of the significance of protein assays laboratories perform this techniques on almost a daily basis. For this experimental procedure, the Biuret and Folin methods were used to determine the protein concentrations of two unknown samples. First the Biuret method (Fig.1) is based on the appearance of peptides bonds in proteins...
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...culture, is the removal of cells from an animal or a plant, and growing the cells in a favourable artificial condition (Wiley, J.M et al, 2013). There are three types of cell culture: primary, this is cultivating the cells away from the parental tissue, secondary, which is when a primary culture is sub-cultured and cell lines, which can be continuous or finite depending on the life span of the culture. Virus titrations are used to estimate the virus concentration; it is a viral quantification technique. When detecting the virus, the cytopathic effect is looked at, whether there is lysis of the cells, vacuolation, formation of syncytia and the presence of inclusion bodies. TCID50, is the measure of the infectious titre. The end point dilution assay quantifies the amount of virus that is required to kill 50% of infected hosts or to produce a cytopathic effect in 50% of inoculated tissue culture cells (Kumar P, 2013). The purpose of the virus titration within tissue culture is to isolate and identify viruses within clinical samples, to carry out research on the viral structure, replication, genetics of the virus and the effect on the host cells, and also to prepare viruses for vaccine production. Results Table 1 Data used to determine the 50% endpoint using the Reed-Muench method Log of virus dilution | Infected test units | Cumulative infected (A) | Cumulative non-infected (B) | Ratio A/(A+B) | Percentage infected (%) | -1 | 5/5 | 37 | 0 | 38/38 | 100 | -2 | 5/5 | 32 | 0...
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...GOLD BUYING PROCEDURE Undocumented/treasure AU Bars PRIOR OR BEFORE IMPLEMENTATION A TABLE MEETING IS A MUST FOR M.O.A SIGNING BY BUYER & SELLER. For sponsored project (Seller is not the one paying the Contingency Fee/ Intermediary/ies paid the fee ) both Parties have agreed that a day before implementation, First Party (SELLER ) will hand over the _____ grams of sliced gold grams from subject gold bars to Second Party ( BUYER ) as “commitment” and to be given to the buyer’s verifier at the venue a day prior ( before ) to implementation as “COMMITMENT” rest in three MAJOR reasons: Assurance gold slice quantity varies as follows : METRO MANILA 100 grams LUZON 200 grams VISAYAS & MINDANAO 300 grams. 1. Avoid last minute back out. ( pag-urong ng seller sa araw ng bilihan / paglabag sa pinirmahang kasunduan ) 2. Avoid switching ( pagpalit ng item na peke ) 3. Serve as key for the Buyer as assurance for expenses incurred ( in caase of seller’s failure ) ( ang sliced grams of gold ay magsilbing pantapal sa nagastos ng BUYER, kung sakali man magkaroon ng hindi pagkatuluyan sa transakayon sa anumang dahilan nilabag ng SELLER sa araw ng buying/implementasyon. ) MIMINUM BUYING PER IMPLEMENTATION 6.2 KG. ( 5 BARS ) 12.5 KG ( 3 BARS ) 74.6 KG. ( 1 BAR ) Transaction/s wherein the Seller or intermeiaries pays for the Contingency Fee for its implementation...
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...The Need for Early Education 2 The Need for Early Childhood Education Early childhood education has many benefits; the most important one teaching young children the necessary learning skills they need to grow socially, and developmentally; children are growing and learning every day so the earlier they start their education the better. The need to provide more children early education is huge, no matter what the family income is. Every child deserves to have a chance to excel in their own future. This paper will address the benefits of early childhood education and the different types of programs available. The positive affect that can happen to children attending early childhood education centers such as head start, pre-k, or a local daycare center are extraordinary. The need for this early education is crucial to children’s futures. “Research shows the benefits to the child include improved readiness to learn, improved early literacy, decreased need for remedial or special education placement, and improved cognitive development. Long term, research shows a stronger likelihood to graduate from high school, improved academic confidence, and more participation in post-secondary education.” (Pennsylvania Economy League, 2009) Children’s early years are so critical. There development, socialization, and language skills are developing and are very important to develop strong. How children develop depends on the skill obtained in there early...
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...Abstract: The case describes in detail the various steps taken by Lenovo, the leading PC manufacturer in China, in its effort to go global. The steps include changing its corporate name from Legend to Lenovo, sponsoring the 2008 Olympic Games and most importantly, acquiring IBM's PC unit. The case examines the need for Lenovo to globalize and critically analyzes the efficacy of the above steps in the company's globalization plans. It also highlights the challenges faced by Lenovo in its path towards globalization. Issues: » How innovation, differentiation and customization can be used as strategic and competitive advantages by a company, to maintain its leadership in the domestic market, and emerge as a global player » The need for globalization and the factors that must be taken into account when a company wants to go global » The measures by which a company can create a global brand » The challenges facing Lenovo in its efforts to go global Introduction In December 2004, the China-based Lenovo Group (Lenovo) announced that it had acquired the personal computer (PC) division of the US-based IT major IBM. Industry analysts termed this as a major milestone for Lenovo in its efforts to globalize its operations. It was also perceived as an important step towards achieving the company's goal of becoming a Fortune 500 company by 2010. According to Liu Chuanzhi (Chuanzhi), the Chairman of the Group, "The purchase will make Lenovo Group the third largest PC...
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...Christianity Essay The religion my group is covering is Christianity. Christianity is one of the biggest and widest spread religions in the world. It was founded by Jesus Christ and his 12 Apostles. Jesus taught his Apostles the word of God and spread the belief that he was the messiah by performing miracles such as healing the sick, feeding thousands with only enough food for one person, and expelling demons from the souls of people he encountered. After Jesus had many followers, people started to worry about the change in the community and the Romans didn’t want to think about what might happen if people started changing their ways so they sentenced Jesus to death. Jesus told his Apostles that he would come back after his death and they should continue to spreading what he had taught them. Three days after Jesus was crucified, he rose from the dead, proving that he was the messiah and everyone should listen to his teachings as they were the true word of God. Christianity is a religion that requires some work if you want a pleasant afterlife, and by this I mean that the only path to Heaven is through enlightenment or being saved. Becoming saved, despite popular belief, is not a public matter, all you have to do is open your heart and except God. God knows what is truly in your heart so you don't have to do it publicly. After you've become saved you can be baptized which is a way of cleansing yourself of sin and basically being reborn as a new person, one who is devoted...
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...CHAPTER 3: METHODOLOGY 3.1 Study Design This research is an experimental study which was conducted to investigate the effectiveness of treatment of Cassia alata as antioxidant effects in cardiovascular system of hyperglycemic rats. The antioxidant activity was tested from the leaf of Cassia alata using its aqueous extract. For this study, 30 Wistar rats weighing between 180 to 200g were used. They were housed in standard cages in a room with a 12 hour light/dark cycle and 50 to 60% relative humidity at a temperature of about 30°C. The animals had fed with standard food and water without limit. This study was conducted in two groups of streptozotocin (STZ) induced diabetic rats in which each group will consist of 15 rats. For the first group of STZ induced diabetic rats had fed with Cassia alata aqueous extract for 20 days meanwhile for the second group of STZ induced diabetic rats had fed with normal saline as a negative control for 20 days. The two groups of STZ induced diabetic had divided into 2 batches. First batch of diabetic rats was consisted of 6 rats for both groups while the second batch was consisted of 9 rats for both groups. All of the rats in two groups had fasted for 12 hour before induced diabetic via STZ injection. After an injection, the rats with level of fasting blood sampling (FBS) above 200 mg/dl will take for further investigation. Henceforth, the treatment was began in which the Cassia alata aqueous extract will administer orally after 48 hour diabetic...
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...4110065A.qxp 9/25/2007 2:39 PM Page 1 ™ Quick Start Bradford Protein Assay Instruction Manual For technical service call your local Bio-Rad office, or in the US, 1-800-4BIORAD (1-800-424-6723) 4110065A.qxp 9/25/2007 2:39 PM Page 5 Table of Contents Section 1 Introduction 1 1.1 Principle 1 1.2 Selecting a Protein Standard 5 1.3 Product Description Section 2 Instructions 9 11 2.1 Standard Assay Protocol 11 2.2 Microassay Protocol 14 Section 3 Data Analysis 18 Section 4 FAQs and Troubleshooting 22 Section 5 Ordering Information 26 Section 6 References 28 Section 7 Appendix 30 4110065A.qxp 9/25/2007 2:39 PM Page 7 Section 1 Introduction The Quick Start Bradford protein assay is a simple and accurate procedure for determining the concentration of protein in solution. It provides ready-to-use convenience by supplying the dye reagent at 1x concentration and two protein assay standards at seven prediluted concentrations. The prediluted standards are conveniently packaged in 2 ml screwcap vials, eliminating wasteful and sharp ampoules, and ensuring protein stability over the shelf life of the product. 1.1 Principle The Bradford assay is a protein determination method that involves the binding of Coomassie 1 4110065A.qxp 9/25/2007 2:39 PM Page 8 Brilliant Blue G-250 dye to proteins (Bradford 1976). The dye exists in three...
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...There are different Instructional System Design models used for the purpose of training that focus on the learning rather than on the teaching... These models as ADDIE guide the process of learning activity and provide the tools that are required for learning process. Instructional Systems Design (ISD) model includes; analysis, design, develop, implement, and evaluations. ISD considered as a formal type used to train health practitioners and others. At this training program for HER implementation, the goal should be determined by assessments and analyses of learners, and then the goal s will be established based on the assessments results. During the training session, different evaluation methods should be established to measure the quality of the training session and the extent of goal achievements. We will train health care practitioners and staff on the new EHR system. We will start with different steps before we begin the training session. It is important to identify all the requirements for training, our goals, training contents, designs the training methods, implementation, and the output of the program. ADDIE Model Phases: Analysis-Assessment-Needs: When-Where-Who. Analyze and assess the learners and the organization’s needs to identify the training session goals that required for this session. Also, at this phase, we determine who and when the training session will occur. Design: To design the training session that meets the learners and trainer learning...
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...have higher peroxidase activity levels because it was under more stress than the plant that was watered with water. Through the use of such techniques as tissue prints, dot-blot assays, electrophoretic analysis, and spectrophotometric assays, peroxidase activity levels were visualized and quantified. The results were as expected; the tissue from the plant that had been treated with Vitamin Water was more purple on the tissue prints and it was spectrophotometrically determined that the plant that had been treated with Vitamin water had a higher amount of peroxidase activity than the one that was treated with water. The results proved that the hypothesis was correct; stress on a plant, Vitamin Water in this case, causes higher levels of peroxidase activity. Introduction: Peroxidases are enzymes that use hydrogen peroxide as an electron acceptor to catalyze oxidative reactions. Peroxidase is believed to play a major role in synthesizing cell walls, preventing toxins and responding to stress. It is responsible for the purple color that is seen throughout the experiments. Peroxidase can be found in different locations in a plant in the form of isoenzymes. Many different experiments can be done to locate, visualize and quantify the activity of peroxidase enzymes. These include, tissue printing, dot-blot assays, electrophoresis, and spectrophotometry. This knowledge of what peroxidase does and how its activity levels can be detected was used to develop...
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...Baghpat Crossing, MEERUT 250 005 UP, India 2 Department of Pharmaceutics, Laureate Institute of Pharmacy, KATHOG 177 101 The. Dehra, Dist. Kangra, HP, India Abstract: Piper betle leaves (Family: Piperaceae) have been used in Chinese and Indian folk medicine for centuries and recently been proposed to be used as a chemopreventive agent because of its anti-oxidant activity. In the present paper, Piper betle leaves were standardized for stomatal index, vein islet and vein termination numbers, palisade ratio, UV fluorescence and different ash values. The Piper betle leaves are earlier reported to possess anticancer potential. Hence, the aqueous extract of the leaves was subjected to cytotoxicity studies on Hep-2 cell line using MTT and SRB assays. The mean CTC50 was found to be 96.25 µg/mL, which proved the potent cytotoxicity and hence, the probable anticancer property of the selected extract. Key words: Betel Leaf Anticancer Phytochemical characterization and its leaves, with a strong pungent and aromatic flavour, are widely consumed as a mouth freshener [4]. The leaves are credited with wound healing, digestive and pancreatic lipase stimulant activities in the traditional medicine [5]. During our exploration of non-toxic and affordable herbal medicinal formulations, the PBL extract and its constituent phenolics were found to show impressive anticancer activities [6-8]. The deep green heart shaped leaves of betel vine are popularly known as Paan in India [9, 10]. MATERIALS AND...
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...cancer [2]. Chemotherapy is restricted by both intrinsic and acquired cell resistance to drugs. [3] This has necessitated the use of natural products for the treatment of cancer. There are compelling evidences for experimental investigations on the efficacy of plant drugs against cancer [4]. It is known that there are links between the inflammatory and nociceptive, oxidative and cancer processes. The ability to inhibit any of the process will definitely lead to the inhibition of the others [5]. Based on the fact that M. calabura fruit possess antioxidant activity, the aim of the present study was to determine the in vitro anticancer activity of methanol extracts of the fruits of M. calabura against HeLa and HEp-2 cancer cell lines using MTT assay. Plant Material and Extraction Collection and authentication of fruits of M. calabura Linn were collected from the surrounding areas of Erode, Tamil Nadu state, India during 2008-2009 and the plant was identified and authenticated and deposited (voucher number: BSI/SC/5/23/09-10/Tech-132) at Tamil Nadu Agricultural...
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...Introduction Ash is a vital ingredient in the canine diet as it is the main constituent of minerals within most diets. Ash content in food is determined by burning the food to calculate the ash percentage left behind. A deficiency in any of the minerals within ash would result in a wide range of health problems which is why it is a legal requirement to display the ash content on food packaging. This experiment is designed to recreate the process undergone by the pet food company to determine the ash content within the food. Method Weigh silica crucible and record weight prior to experiment. Grind over 5g of food sample in pestle and mortar and measure 5g of ground sample into the crucible. (W0) Place the dish in a baking oven for 24hrs to dry out the sample. Reweigh the sample in silica crucible, This is now the true weight of the sample (W1) Set up the bunsen burner on top of the heatproof mat with tripod and clay pipe triangle above to support the crucible. Ignite the bunsen burner and begin to heat the sample slowly ensuring the sample does not set on fire as this will result in a loss of material to the atmosphere in the form of particles. The sample should turn black. Once the sample reaches roughly 100°C continue to heat to 500°C until the sample turns white. Allow sample to cool to room temperature in a dessicator. Weigh the finished sample (W2) Calculate the ash content (X): (W2-W0)/(W1/W0)x100=X Materials: Silica dish Baking Oven Bunsen Burner ...
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...of the lab gives you a tube of enzyme and tells you the following before disappearing into the office to write more grant proposals: ➢ There is 50 µl of enzyme stock solution. The enzyme is expensive to purify, so follow the protocol exactly, using as little of the stock solution as possible. ➢ The concentration of the stock solution is currently not known, but a 1 mg/ml concentration of the pure enzyme has an A280 nm of 2.0. ➢ You’ll be performing the assay on 12 samples. ➢ Make enough of each working solution so that you have at least 400 ul to work with when you do the assay (to cover any waste and/or inefficiencies in pippetting). Using the spectrophotometer to read the absorbance at 280 nm, you get a reading of 0.784. 1. (2 pts) What is the concentration of the solution in the cuvette? What is the concentration of the stock solution? (Show your work, box your answers.) 2. (1 pt) Calculate the minimum amount of each working solution would you need in order to do the number of assays requested. How much excess has the PI instructed you to make? 3. (6 pts) Figure out...
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...BÀI KIỂM TRA Ngày 28/8/2014 Họ và tên: …………………………………………… Bài 1: Từ vựng 1. 2. Jing lỉ hảo! (chinh lỉ hảo):…………… 3. Wan an (uản an):…………………….. 4. Dùi bù qỉ (tuầy pù chỉ):………………. 5. 35:……………………………………. 6. 201:…………………………………… 7. Nu hái (nủy hái):……………………… 8. Hái hảo (hái hảo):…………………….. 9. Jie hun (chiế huân):…………………… 10. Gong rén (cung rấn):………………….. 11. Duo jiu (tua chiểu):……………………. 12. Yao`:…………………………………... 13. Xiang’(xiẻng):…………………………. 14. Zhi dào (trư tào):………………………. 15. Rén (rấn):………………………………. 16. Xỉao shi (xỉeo sứ):……………………... 17. Dian (tẻn):……………………………. 18. Mei tian (mẩy thiên):…………………. 19. Bái ban (pái pan):…………………….. 20. Ké bù ké ỷi (khứa bù khứa ỉ):………… 21. Jin tian (chin thiên):…………………… 22. Liang ge ỳue (liengr cưa yuề):………… 23. Yi yue (I yuề):…………………………. 24. Dà (tà):………………………………… 25. Gao xìng (cao xình):…………………… 26. Qỉng wèn (chỉnh uần):…………………. 27. Bái sè (pái sừa):……………………….. 28. Huang se (hoáng sừa):…………………. 29. Nả lỉ (ná lỉ):……………………………. 30. Kuàng gong(khoàng cung):……………. 31. Gong chảng (cung chảng):…………….. Bài 2: Điền từ vào chỗ trống: 1. Ủa chieo`……………………………………………… 2. Chỉnh……………..…ràng ủa dừ chỉ……………………..í xìa. 3. Ủa cha yẩu…………………cưa rấn. 4. Ỉ chén ủa sừ……………………………………. 5. Ủa xỉ hoan………………………………..sừa. Khoanh...
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