Biochemistry is the study of the chemical processes that happen in the cells of humans, animals, plants and other living creatures. Biochemical protocols refers to procedures used in a biochemical laboratory, which can be involved in the production of new drugs, scientific research or medical analysis of samples, such as blood and urine. Biochemical protocols include cell staining, antiprotozoal activity, lymphocyte migration and electrophoresis.
Cell Staining Protocol
The use of dyes helps in the identification of internal cellular structures, when observing samples under a microscope. Trypan blue is an acidic dye only used to analyze dead cells, because when still functioning, cells do not absorb the substance. During this protocol, technicians wash the sample with PBS or phosphate buffer solution before adding trypan blue.
Antiprotozoal Activity Protocol
Protists are microscopic organisms, some of which are related to serious diseases, such as malaria and trypanosomiasis. Phenethyl-4-aminopiperidine is a group of chemical compounds that present antiprotozoal activity. In this protocol, technicians cultivate protists in a medium containing horse blood serum, amino acids and glucose, as well as the chemical compounds 2-mercaptoethanol and sodium pyruvate. Later, phenethyl-4-aminopiperidines are added to samples to test the protists' resistance to the drug.
Lymphocyte Migration Protocol
Lymphocytes are immunodefense cells that form the blood. These cells can move out of the blood stream, to capture bacteria and other pathogens. Scientists can study these cells by using a fluorescent dye called r CFSE or carboxyfluorescein succinimidyl ester.First, scientists prepare a cell suspension with 20 million cells per-milliliter in FBS or fetal bovine serum and PBS buffer. Then, CFSE is added. The suspension rests for eight minutes, before warmed FBS is also added. Cells are incubated in a 98.6 degrees-Fahrenheit water bath for 10 minutes, before being finally centrifuged and washed three consecutive times in a solution of FBS/PBS.
Tris-tricine Electrophoresis Protocol
Tris-tricine is a chemical compound used as buffer solution to balance the pH of a solution. It is also used as a gel in electrophoresis protocols, which refer to the analysis of proteins and other cellular components by their movement in an electric field. To prepare tris-tricine gel for electrophoresis protocols, scientists make a mixture of double distilled water, acrylamide, chlorine, tris-tricine and sodium lauryl sulfate (SDS). Later, glycerol, ammonium persulfate and tetramethylethylenediamine are added.
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