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Transfection and Immunofluorescence Staining
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Transfection and Immunofluorescence Staining
LacZ plasmid reporter and the GFP are used as controls to assess the efficiency of the transfection procedure. Within twenty four to forty-eight hours of transfection, expressions of plasmids should be evident. Positive control can be co-transfected with plasmids of interest or maintained in a different well.
To corroborate the specificity of primary Antibody binding, and to cancel out cellular interactions of proteins, a staining control is very relevant. Isotype Antibodies are used to because they retain characteristics that are nonspecific and have no target cells’ specificity.
Alternatively, phalloidin can be used for staining in place of the Antibodies for the staining. In cells, both nonmuscle and muscle, labeled conjugates of phalloidin tend to bind in a ratio that is stoichiometric; a phallotoxin per every unit of actin, because of their similar affinity for minute and large filaments. But unlike the Antibodies’ action against actin, the phalloidin do not attach to the monomeric G-actin.
Advantages of the GFP
The Green Fluorescent Protein (GFP) has assumed a central role as a tool in visualization of both temporal and spartial patterns. There are two major reasons for this. The first is that it helps avoid artifacts because the samples do not need to be fixed to infuse the membrane to permit the Antibodies’ diffusion.
Unstable variants are used in the time dynamics determination of protein expressions, but stable variants are used in the location of specific structures. GFP has variants with stability differences ranging from relatively low to rather very high thus is important in both the cases.
DAPI, as a nucleic acid stain, preferentially stains the dsDNA. With little cytoplasmic labeling, DAPi specifically stains the nuclei. When DAPI staining has been combined with primary and secondary antibodies of choice, signals are amplified, and the choice of isotypes are increased (up to five) to be simultaneously distinguished.
Reference
In Kabelitz, D., & In Kaufmann, S. H. E. (2010). Immunology of infection. Amsterdam: Elsevier.
The Biasness of the Healthcare System during the Covid-19
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The Biasness of the Healthcare System during the Covid-19 Pandemic
The covid-19 pandemic shook the world when it first hit in 2019. It proved to be more than a health crisis as it disturbed all country sectors and the global economy. The pandemic also exposed some of the government’s weaknesses, especially the health department. The political cartoon represents how the healthcare system is biased against certain groups of people in the community, especially during the pandemic. It terms them as ‘underlying conditions.’ according to the cartoon, the most affected groups were the poor, black people and nursing home residents during the pandemic, yet the healthcare system downplayed it. The cartoon approaches the issue of biases in the health system using an interviewer interviewing a healthcare operator.
The cartoonist tries to create an emotional appeal by introducing a hospital bed and labeling the names of the disadvantaged groups on it. This picture is meant to capture the reader’s attention and persuade them to connect their emotions and relate to the discrimination that heightened during the pandemic against these groups of people. It stirs pity and empathy towards the poor, black people and nursing home residents. With logos, reason was achieved through the healthcare system trying to hide the unfairness that black people, the poor and the nursing home residents by generalizing the issue. The image depicts the social problem that has been in existence for a long time and was felt even more during the pandemic. In the picture, the healthcare uses the ‘underlying conditions’ to represent being poor, black or a nursing home resident, which is unfair and inconsiderate.
Social interests are guidelines and attributes that contribute
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Social interests are guidelines and attributes that contribute to an individual’s sense of belonging. It constitutes factors like participating in societal activities, cooperating, empathy, respect, and encouraging other community members. A person’s interests should align with the societal interest for a peaceful and comfortable society to exist. Challenges can occur if a person’s interests don’t align with societal interests.
An individual may lack a sense of belonging. An individual is likely to lack a sense of belonging if the interest of their society is quite different from theirs. It will be difficult for them to relate to some of the activities that go on in society. Moreover, it limits them from socializing and expressing themselves freely for fear of facing judgment from other society members. It can be difficult for them to cope and exist peacefully, which is the whole meaning of being part of a community. For this reason, they are not able to exploit their full potential.
The collide of personal interests and social interests can result in stagnating community growth. Individuals might lack interest in participating in communal activities if their interests do not align with societal values. For a community to flourish, it requires all its members to be part of various projects developed in the community. If part of the community doesn’t share its interest, completing these projects won’t be easy. Hence stagnation is experienced in the community.
Furthermore, individuals in society will feel the need to isolate themselves from a community that does not share their interests. It will result in individualism since people only care about their personal needs and don’t look out for their neighbors. The sense of a community will thus disappear. Therefore, people need to be part of a community they identify with in terms of their interests. It will make them comfortable and contribute to the growth of the community at large.