University Presentation Showcase: Undergraduate Division
Detecting Phosphorylated Proteins by Performing Redox Proteomics
Presenter Hometown
Lexington
Major
Nursing
Department
Chemistry
Degree
Undergraduate
Mentor
Tanea Reed
Mentor Department
Chemistry
Recommended Citation
Grasso, Maya M., "Detecting Phosphorylated Proteins by Performing Redox Proteomics" (2024). University Presentation Showcase Event. 28.
https://encompass.eku.edu/swps/2024/undergraduate/28
Abstract
Traumatic Brain Injury (TBI) is defined as an event that occurs when trauma causes damage to the brain. Damage occurs due to the primary injury (impact due to initial event) and secondary injury (neuroinflammation causing phosphorylation in neuronal cells). For normal cellular function to occur phosphorylation and dephosphorylation must be undergone in molecules. The effects of phosphorylation and dephosphorylation on different proteins in the brain due to Traumatic Brain Injury and during recovery of TBI not understood and lack research. To analyze phosphorylated proteins, 2D gel electrophoresis was performed. Samples from rat brains were dissolved in IPG rehydration buffer prior to isoelectric focusing electrophoresis (IEF). IPG strips were rehydrated with either 200ug or 600ug of protein sample dependent on type of stain used. Isoelectric focusing was then performed using a horizontal electrophoresis system for 24 hours. Strips were then removed from the mineral oil and incubated in 10 ml of two equilibration buffers for 15 minutes each. Gel electrophoresis was then performed at 200V for 65 minutes. Different staining techniques used to include Pro Q diamond, Quercetin, Coomassie Blue and Sypro Ruby stain following protocol by Wang et al. The result of this experiment determines that the protocol identifies protein samples stained with ProQ diamond, Quercetin, Coomassie blue, and Sypro Ruby which can be evaluated through image results. In an effort to better understand the phosphorylation process of proteins involved in traumatic brain injury, 2 SDS-PAGE be assessed.
Presentation format
Poster
Detecting Phosphorylated Proteins by Performing Redox Proteomics
Traumatic Brain Injury (TBI) is defined as an event that occurs when trauma causes damage to the brain. Damage occurs due to the primary injury (impact due to initial event) and secondary injury (neuroinflammation causing phosphorylation in neuronal cells). For normal cellular function to occur phosphorylation and dephosphorylation must be undergone in molecules. The effects of phosphorylation and dephosphorylation on different proteins in the brain due to Traumatic Brain Injury and during recovery of TBI not understood and lack research. To analyze phosphorylated proteins, 2D gel electrophoresis was performed. Samples from rat brains were dissolved in IPG rehydration buffer prior to isoelectric focusing electrophoresis (IEF). IPG strips were rehydrated with either 200ug or 600ug of protein sample dependent on type of stain used. Isoelectric focusing was then performed using a horizontal electrophoresis system for 24 hours. Strips were then removed from the mineral oil and incubated in 10 ml of two equilibration buffers for 15 minutes each. Gel electrophoresis was then performed at 200V for 65 minutes. Different staining techniques used to include Pro Q diamond, Quercetin, Coomassie Blue and Sypro Ruby stain following protocol by Wang et al. The result of this experiment determines that the protocol identifies protein samples stained with ProQ diamond, Quercetin, Coomassie blue, and Sypro Ruby which can be evaluated through image results. In an effort to better understand the phosphorylation process of proteins involved in traumatic brain injury, 2 SDS-PAGE be assessed.
