Title: Does Basigin-2 Bind to Basigin on Monocytes to Generate an Inflammatory Response? 1) Introduction a) Overview of Inflammatory and immunologic response. i) Differentiate and describe between Innate Immunity and adaptive immunity. ii) Differentiate and describe between monocytes and macrophages (Innate Immunity) and B cells defining antibodies and T cells defining cytokines (adaptive immunity). iii) Describe the importance of receptors for innate immunity. iv) How does TLR4 and Basigin along with Basigin-2, which is retina-specific described as receptors induce innate immunity? v) Redzi et. al 2011 portrayed that Basigin-2 activates 1L-6 which is a pro-inflammatory cytokine in monocytes using TLR4. 2) Purpose i) To portray evidence …show more content…
3) Materials and Methods i) Raw cells plated in 4 chamber slides each slide containing 8 mini cube wells. ii) Four percent of paraformaldehyde was added to all chamber slides. iii) The paraformaldehyde was removed and all the wells in each chamber slides were washed 3 times with PBS. iv) A pre-incubation buffer was added to each chamber slide and incubated at 37 C for 30 minutes. v) After incubation, primary antibodies Basigin and TLR4 were added to 2 chamber slides of each antibody respectively. vi) All 4 chambers with respective primary antibodies were incubated at 37 C for 30 minutes and then stored in the refrigerator. vii) All the wells in each chamber slides were washed 3 times with PBS. viii) Secondary antibody, Alexa 488-GAR was then added to all 4 chamber slides and the chambers slides were incubated at 37 C for 30 minutes. ix) The secondary was then removed and all the wells in each chamber slides were washed 3 times with PBS. x) The wells were then covered with cover slips and stored in the refrigerator. xi) The cells were then fixed and stained. xii) Basigin-2 and 6xHistidine-tag were then added to the cells. xiii) The cells were then fixed and
The procedure of the lab on day one was to get a ring stand and clamp, then put the substance in the test tube. Then put the test tube in the clamp and then get a Bunsen burner. After that put the Bunsen burner underneath the test tube to heat it. The procedure of the lab for day two was almost exactly the same, except the substances that were used were different. The
The immune system is made up of a network of cells, tissues, and organs that work together to protect the body, and it defends the body from “foreign invaders.” Immunity can be divided in two three different defenses, and these are defined as first, second and third lines of defense. The first line of defense for the immune system is the primary defense against pathogens entering the body from the surface in order to prevent the start of disease and infection. Some examples of the first line of defense is the skin, protecting the external boundaries of the body, and the mucous membranes, protecting the internal boundaries of the body. Although the skin and mucous membranes work on the internal and external boundaries, they both release chemicals
“Immune Response: MedlinePlus Medical Encyclopedia.” National Library of Medicine - National Institutes of Health. Web. 18 Dec. 2011. .
One disadvantage of the single antibody system is its reduced sensitivity as the signal is not ...
After ten minutes had passed, I collected the ingredients needed to perform a gram stain. I got the primary stain, crystal violet, and flooded my smear for sixty seconds, and then rinsed the color off with water until the water ran clear. I then flooded the smear with the mordant, grams of iodine, and let that sit on the slide for sixty seconds as well. I then rinsed the grams of iodine off with water and applied alcohol to the smear to decolorize the cells; however I made sure not to over decolorize and only put enough drops on the smear till the purple ran clear. I then rinsed the slide with water and flooded the smear with safranin, the counter stain and let it sit for sixty seconds and then rinsed the color off with water.
Immunohistochemistry is a technique that involves the use of antibody-antigen interactions in order to identify cellular and tissue constituents. One way this can be done by labelling known antibodies with enzymes which produce a coloured product after reacting and then monitoring the sample to see whether a reaction has taken place. In immunohistochemistry, the preservation of the antigenic determinants, also known as epitopes, and binding sites is vital in order to ensure that an accurate result is obtained. However, tissue samples need to go through various processes in a pathology laboratory before they are ready for testing and some of these processes can alter the structure of the epitopes so it’s important to consider this when choosing reagents in order to reduce the risk of this happening.
Seattle Education Project. (2013, November). STEP: The Immune System - An Overview, [Online]. Available: http://www.thebody.com/step/immune.html [11/12/14].
Experiment: First prepared a well plate with the appropriate amounts of distilled water, HCl, and Na2S2O3 in each well according to the lab manual. The well where the reaction
Non neutralizing antibodies interfering with protective response of the broadly neutralizing antibodies (bnAbs). Immune correlates of the RV144 trial have shown production of Non neutralizing antibodies
...d the absence of IgE binding from patients allergic to protein from the source as well as histamine release from basophiles is required to be carried out.
== § Test tubes X 11 § 0.10 molar dm -3 Copper (II) Sulphate solution § distilled water § egg albumen from 3 eggs. § Syringe X 12 § colorimeter § tripod § 100ml beaker § Bunsen burner § test tube holder § safety glasses § gloves § test tube pen § test tube method = == = =
Inflammation which is part of the innate immune system is a process by which the body reacts to injury protecting it from infection and foreign substances with the help of the body’s white blood cells “Inflammation can be defined as the body’s local vascular and cellular response to injury caused by factors that invade and injure the body from the outside (exogenous factors) or factors within the body that result in cellular or tissue injury (endogenous) factors” (Battle, 2009, P 238). Factors such as bacteria, viruses, burns, frostbite, chemical irritants, immune reactions and physical injury are examples of factors that can cause inflammation through different mechanisms. It is a protective mechanism with rapid response that neutralizes or destroys agents that causes injury and creates a barrier that limit the injury and prevents its spread to normal tissues (Battle, 2009). Also, it has elements that removes debris and heals the wound generated by the injury. It can be divided into acute and chronic inflammation.
Create wells: put a comb template in the middle of the tray; wait until the mixture becomes solid. After, remove the comb standing straight. 4. Remove rubber ends: transfer the gel tray into the horizontal electrophoresis and fill it with the concentrated electrophoresis buffer. 5. Materials and methods: Experiment: 1st, prepared milk samples should be already done by the teacher.
Adaptive immune system happens much quicker to the presence of an “infection creating potent mechanisms for neutralizing or eliminating the microbes. There are two types of adaptive immune responses: humeral immunity, mediated by antibodies produced by B lymphocytes, and cell-mediated immunity, mediated by T lymphocytes.”
Added one of the 1000 gram dry samples to the top of spiral where water is coming out of the distributor.