Write up - using a double-headed needle We will inject 1mL...

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Mayra Trujillo 9/14/2015 During this laboratory period, we will induce a humoral response into 8 out of 10 mice with the antigen bovine serum albumin (BSA) and a TBA protein. We will determine the levels of specific antibodies (anti- BSA) & (anti-TBA protein) through the use of an enzyme linked immunosorbent assay (ELISA). Also, we will be working with Complete Freund’s Adjuvant (CFA). It is extremely important to use gloves and protective eyewear when handling CFA. The goal of this experiment is to answer the following question: Does the immune response vary if it responds to two antigen instead of one antigen. The mice will be divided into 4 groups. The first group will composed of three mice & will be injected with BSA, TBA protein, & CFA. The second group of mice will be composed of 3 mice & will be injected with TBA protein & CFA. The third group of mice will be composed of 2 mice & will be injected with BSA & CFA. The fourth group will serve as the control group and will not be injected. The emultion will be created by using equal parts (0.25 to 0.5mg/ml) of the antigen(s) and CFA
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Unformatted text preview: using a double-headed needle. We will inject 1mL of the emul±on using a syringe F³ed with a 25 G needle into the peritoneal cavity of the mouse. A´er two weeks the intraperitoneal injec±on will be repeated using Incomplete ²reud’s adjuvant. We will then kill the mouse using CO2 asphyxia±on and withdraw blood from the heart. Using a syringe F³ed with 23G needle we will withdraw slowly from the apex of the heart. Transfer blood into a micro centrifuge tube. Es±mate volume and place small wooden rod into the tube & allow the blood to clot at RT incuba±on followed by overnight 4 °C . Remove clot carefully and centrifuge the tube. Transfer serum into a new tube and es±mate volume. We will determine the levels of speciFc an±bodies using ELISA. We will then compare all four groups and analyze the levels of speciFc an±bodies in all four groups to determine the answer to the ques±on stated above....
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