Ataxia-telangiectasia (A-T) or Louis-Bar syndrome is a rare, autosomal recessive, inherited disease that affects different systems such as the immune and nervous systems (2). The name of the disease provides insight to what happens when an individual has this. Ataxia means poor coordination while telangiectasia mean enlarged blood vessels (2). Individuals show symptoms of this disorder at an early age, typically before the age of 5. Since the immune system is compromised, individuals are at high risk for infections which can lead into death for most patients. Cancer is also another cause of death because of the protein ATM is not able to function properly (1,2,4,5).
The ATM gene provides a manuscript to making a protein that regulates cell …show more content…
Both samples will be used to compare to each and to determine what had caused the tumor. We will be immunoblotting (western blot) for the ATM protein (3). The tumors will be isolated and grown in culture so we have enough samples to work with (1). We prepare the samples for two-dimensional gel electrophoresis using 1D/2D Electrophoresis ZOOM IPGRunner System[1,3]. This process allows the proteins to be separated and particularly were looking for the ATM protein (1). The proteins are separated by their natural charges and dissolved in a solution of uncharged detergent with a combination of β-mercaptoethanol and the denaturing reagent urea (1). The solution breaks down the polypeptides chains but leaving their natural charges intact (1). These broken down polypeptide chains undergo a process called isoelectric focusing, a process where proteins in a pH gradient are separated based on their isoelectric point (1). The isoelectric point is the pH level where the protein does not have a net charge anymore (1). All this constitutes the first dimension of the two-dimensional polyacrylamide-gel electrophoresis. The second process consists of another electrophoresis but at a right angle to the direction to the first electrophoresis that was done (1). SDS is added in this second process and separates the proteins based on their size forming the one-dimensional SDS-PAGE (1). The first gel is soaked in SDS and situated on the edge of the SDS polyacrylamide-gel slab where the polypeptide chains move towards to and form a spot (1). Thus the second-dimension of two-dimensional polyacrylamide-gel electrophoresis is completed (1). Now we can distinguish the proteins that are in A-T diagnosed patients and those who do not have A-T. Approximately 90% of A-T patients do not have detectable ATM protein and only
The ATM gene provides a manuscript to making a protein that regulates cell …show more content…
Both samples will be used to compare to each and to determine what had caused the tumor. We will be immunoblotting (western blot) for the ATM protein (3). The tumors will be isolated and grown in culture so we have enough samples to work with (1). We prepare the samples for two-dimensional gel electrophoresis using 1D/2D Electrophoresis ZOOM IPGRunner System[1,3]. This process allows the proteins to be separated and particularly were looking for the ATM protein (1). The proteins are separated by their natural charges and dissolved in a solution of uncharged detergent with a combination of β-mercaptoethanol and the denaturing reagent urea (1). The solution breaks down the polypeptides chains but leaving their natural charges intact (1). These broken down polypeptide chains undergo a process called isoelectric focusing, a process where proteins in a pH gradient are separated based on their isoelectric point (1). The isoelectric point is the pH level where the protein does not have a net charge anymore (1). All this constitutes the first dimension of the two-dimensional polyacrylamide-gel electrophoresis. The second process consists of another electrophoresis but at a right angle to the direction to the first electrophoresis that was done (1). SDS is added in this second process and separates the proteins based on their size forming the one-dimensional SDS-PAGE (1). The first gel is soaked in SDS and situated on the edge of the SDS polyacrylamide-gel slab where the polypeptide chains move towards to and form a spot (1). Thus the second-dimension of two-dimensional polyacrylamide-gel electrophoresis is completed (1). Now we can distinguish the proteins that are in A-T diagnosed patients and those who do not have A-T. Approximately 90% of A-T patients do not have detectable ATM protein and only