Cell Observation by Fractionation

Cell Observation by Fractionation Cell fractionation techniques are used to separate cells into their cell organelles so that the function of each part can be observed in more detail. The main tools used are centrifuges and test tubes which are paired and rotated at a certain speed which has a speed according to needs. A centrifuge with a speed of up to 80,000 revolutions per minute (rpm) is called an ultracentrifuge so it exerts a force on the particles of up to 500,000 times the force of gravity (500,000 g). Fractionation begins with homogenization, to disrupt the cells, that is, to break the cells without damaging the organelles. Treatment with a centrifuge will separate the cell parts into two fractions, 1) pellets, which are larger structures that collect at the bottom of the test tube. 2) supernatant, which consists of smaller cell parts suspended in the liquid above the pellet. The supernatant was poured into another tube and centrifuged again. This process continues to be repeated with increasing speed at each stage, thereby collecting increasingly smaller components in successive pellets. Fractionation with homogenization, as intended for purification, can also be applied to determine certain types of proteins as products of protein synthesis. To purify a target protein, the procedure can be arranged using gradual steps to reach the intended target protein. The fractionation process begins with homogenization of the tissue placed in a test tube and then centrifuged so that the supernatant (the top of the solution containing light molecular weight materials or organelles) and the pellet (the bottom of the tube which is rich in cell nuclei and cell debris) is separated. which has a large molecular weight). The supernatant is transferred back into another test tube and centrifuged again as needed, so that after doing this several times the result will be a pellet rich in ribosomes (as protein producing organelles). The results obtained are then given other treatment or analyzed as needed. Protein purification is needed to study how chemical reactions are catalyzed or what other functions they perform as well as their regulatory or regulating activities. Many enzymes are composed of many protein chains where one chain will act as a catalyst for a chemical reaction and the other chain will regulate when and where the reaction will take place. By separating each of these proteins or their activities, they can be analyzed. For example, purification and ways to study the activity and composition of DNA replication will prove the working mechanism for DNA replication during cell division. The shape of the fold, three-dimensional structure, or conformation of a protein are vital conditions related to the function of the protein. To understand the relationship between protein function and structure, you need to know the detailed structure of the protein. The method that can be used to determine the structure of protein, DNA and RNA complexes is the x-ray crystallography method. With the help of computer analysis, it will be possible to know the location of each atom, molecular complex and then determine its type.