The Isolation‚ purification and identification of Proteins assayed From Bovine Liver Using SDS Gel Electrophoresis‚ Mass Spectroscopy and Western Blotting Abstract The purpose of the experiment was to isolate and recognize varying protein solubilization and assaying methods by using bovine liver protein. The experiment implicated the impact of different types of solvents like ethanol‚ water‚ PBS‚ PBS+1% Triton x-100‚ and PBS+2% SDS on protein solubilization. Bradford and Ghosh/Dumbroff
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INTRODUCTION A major obstacle in the successful treatment of cancer is the development of resistance mechanisms to drug treatment. Various cellular changes that have been implicated in the development of drug resistance in cancer cells include: the increased expressions of P-glycoprotein (a multidrug transport protein) and multidrug resistance-associated protein (MRP-1); increased levels of the cellular detoxification protein‚ glutathione; and changes in the expression of apoptosis associated
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Review Sheet Exercise 12 Serological Testing NAME Kali Rothfuss LAB DATE/TIME 6/22/11 Positive and Negative Controls 1. Why are there a number of washing steps in serological tests? The are a number of steps needed in order to remove any non specific binding that may have occurred. 2. Describe how you would know that you had a “false positive” result. What does this mean for the rest of your results? A positive result with a negative control indicates a “false positive”
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The samples were boiled for 3 minutes and centrifuged for 20 seconds. A polyacrylamide gel was assembled and the samples were loaded into it. The gel ran for approximately 45 minutes at 160 V. A western blot analysis was done in order to examine the Erk1/2 phosphorylation of the NIH3T3 cells. Three pieces of Whatman paper were wet in transfer buffer and placed on the anode plate. A wet PVDF membrane was placed on top of the Whatman paper. The gel was removed from holder and the stacking gel was removed
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2.4. Animals One hundred twelve adult male Wistar rats (Pasteur Institute‚ Tehran‚ Iran) weighing 220-250 g were housed in groups of four per cage in a controlled environment (temperature 23±1°C‚ 12h light/dark cycle) with free access to food. Rats were habituated to the new environment for seven days before the test. All experiments were performed in accordance with the guide for the care and use of laboratory animals (IR.SBMU.nrc.REC.1390.20) approved by the Research and Ethics Committee of the
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Chicken. In order to able to detect and analyze proteins based on their ability to bind to a specific antibody‚ the SDS-PAGE and Western Blot was performed. SDS Polyacrylamide Gel Electrophoresis (SDS-PAGE) is a very common technique used to separate proteins based on molecular weight under the influence of an applied electrical field and then used to prepare for the Western Blot (#1 Lehninger). The support medium used is a polyacrylamide gel and‚ it also used sodium
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REVIEW SHEET EXERCISE 12 Serological Testing NAME: LAB TIME/DATE: Positive and Negative Controls 1. Why are there a number of washing steps in serological tests? They are needed in order to remove any non specific binding that may happen. 2. Describe how you would know that you had a “false positive” result. What does this mean for the rest of your results? If it has a negative control then it’s a false positive so the results are invalidated. 3. Describe how you would
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1. What are antibodies and why are antibodies ideal for targeting? An antibody‚ also known as an immunoglobulin‚ is a large Y-shaped protein used by the immune system to identify and neutralize foreign objects such as bacteria and viruses. The antibody recognizes a unique part of the foreign target‚ termed an antigen.[1][2] Each tip of the "Y" of an antibody contains a paratope (a structure analogous to a lock) that is specific for one particular epitope (similarly analogous to a key) on an antigen
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Materials and Methods Growing the G Strain and Preparing the GCE (rGFP Crude Extract): To grow the bacterial culture‚ use 10 ml of liquid LB growth media for incubation. 500 ml of the bacterial culture is allowed to grow overnight at 37°C. It is later shaken vigorously to increase the OD600 to 0.5‚ which means that time equals zero. At time zero‚ 1 mL of the culture is transferred into a 1.5 mL centrifuge tube and centrifuged for 5-10 minutes to obtain a pellet. The supernatant should be discarded
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Audit Proposal Do Private Laboratories provide HIV/AIDS Counselling and Testing? Nigeria as a country has a population of 167 million people (1) while her HIV prevalence rate is 3.4 %; an approximately 3.4million people are living with HIV/AIDS. This number represents the second highest HIV burden in the world second only to South Africa (2). The Government of Nigeria has intervened and put in place different measures to reduce this incidence to its barest minimum amongst which is the National
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