Immunolocalization of the microtubule cytoskeleton Essay

Immunolocalization of the microtubule cytoskeleton - Essay Example The most prominent methods are: Western blot, spectrophotometry, enzyme assay, immunoprecipitation and immunostaining. In immunostaining, the method used during this procedure, an antibody is used to detect a specific protein epitope. These antibodies can be monoclonal or polyclonal. Then enzymes such as horseradish peroxidase or alkaline phosphatase are commonly used to catalyse reactions that give a coloured or chemiluminescent product. Fluorescent molecules can be visualised using fluoresence microscopy. During this experiment, the distribution of the protein tubulin in normal rat kidney (NRK) cells is explored. A monoclonal antibody that is specific for the -subunit of tubulin is used. Tubulin polymerizes into long, 25-nm wide microtubules that we will visualize with tubulin antibodies. The formation and maintenance of microtubules is cold sensitive. At 4C, microtubules are destabilized and they depolymerize. At 37C, human body temperature, they remain polymerized. Photograph 1 shows cells incubated at 4 0C, while photograph 2 shows cells incubated at 37 0C. At this stage of the procedure, microtubules cannot be detected in either of the photographs. 3 separate plates are used to create the NRK cells culture. ... The coverslip is then immersed in 3.7% formaldehyde (in PBS) at room temperature. Immersion in 0.2% Triton-X 100 (in PBS) detergent at room temperature follows. To prevent the subsequent antibody from sticking non-specifically to the cells on the coverslip, the coverslips should be incubated with cell-side-up with a 100-l droplet of 3% (w/v) bovine serum albumin (BSA). The incubation is done on a piece of Parafilm in a humid Petri dish for 15 minutes. An anti-tubulin dilution is prepared with 1 part antibody + 75 parts 1% (w/v) BSA in PBS. A 200 l total of antibody is necessary for the three coverslips. This tubulin antibody is a monoclonal antibody (12G10) and recognizes the -subunit of tubulin. Then each coverslip is overlayed each with a 100-l droplet of the diluted tubulin antibody. After that, a secondary antibody with 1 + 200 in 1% (w/v) BSA in PBS is used. The coverslips are placed back into the Petri dish chamber and overlay with a 100-l droplet of secondary antibody and they are incubate for 30 minutes at 37C. Therefore, we have 3 coverslips, all treated with the secondary antibody, 2 of them are treated with the primary antibody (one of them is incubated at 37C, and the other incubated at 4C) and 1 that is not treated with the primary antibody (incubated at 37C). The coverslips then are placed back into the Petri dish chamber and are overlaid for 30 minutes at 37C with a100-l droplet of Vectastain ABC reagent. This reagent contains a complex of avidin and the enzyme horseradish peroxidase (HRP). They are washed again with PBS. The drops of DAB (diaminobenzidine; a carcinogenic compound) are applied to the coverslips and incubate at room temperature for 5 minutes. After DAB incubation,

Cultural Competency in Health Promotion Essay Example | Topics and Well Written Essays - 750 words

Cultural Competency in Health Promotion - Essay Example In this way, communication during health care is limited because the patient has no obligation to communicate his/her feelings. Lack of health education in our culture has led to poor medical care hence the inadequacy in the knowledge about the health services being offered. Some cultural groups have a poor decision leader who must be available for a decision to be arrived at. To overcome the above problems, there must be a development of culturally competent practices that involve self analysis and change of attitude in various cultures. Health care providers should expand their knowledge and not only focus on culture differences while striving to understand the cultural dynamics affecting medical practice and healthcare in general. They should learn about cultures they sever through, they ought to know patients assessment techniques, readings and community activities. Medical practitioners should improve on the cross-cultural communication by being aware of the difference in social norms and knowledge of different languages and lifestyles. The use of assessing health literacy, interpreters and the bilingual patient education materials in educating the community on health matters (Spector, 2009) can play an important role especially from the training period of the medics. Involvement of the community in health matters through health fairs, ethnic festival participation and constant communication with cultural decision makers. These leaders help in providing cultural competent care by strengthening communication ties. It is important for nursing practitioners to work towards cultural competency through provision of culturally competent patient care. Nurses should therefore, be sensitive and open-minded to avoid being compromised by the cultural differences in perception of illness. While communication skills should be used in the best interest of the patient, limits must be set to ensure that the patient does not manipulate the treatment required. Nurses can de velop cultural competency by understanding individual and organizational views of different cultures. Acquisition of background knowledge about a patient on the cultural views can help in decision making. Having collective communication skills will then enable one to get the real meaning of words even if the patient is from a different culture. There should be strong ties between the nurse and the community. However, nurses must be aware of national and professional priorities for caring of cross culturally populations. Responses from 3 families from different cultures The three families interviewed were from middle-class status of life. The families interviewed were an African family, a Latino family, and a white American family. The interviews were contacted at the community hospital. It was to understand their take on cultural differences in medication and modern treatment. Health maintenance The African family did not have better ways to maintain their health compared to other f amilies. They only look for health services when they are sick. It was the responsibility of the family to take the sick person to the hospital when other alternatives have failed. Latinos families have an average medical attention towards maintaining their health. They rely very much on health centers for the services. The white American fa