Creating an N-Methyl-D-Aspartate Receptor 2B-Specific Vector with Green Fluorescent Protein and Optimizing a Transfection Protocol
Session Number
Q19
Advisor(s)
Amanda Gross, Northwestern University Roger Kroes, Northwestern University Joseph Moskal, Northwestern University Mary Schmidt, Northwestern University
Location
B-131 Grainger
Start Date
28-4-2016 8:25 AM
End Date
28-4-2016 8:50 AM
Abstract
N-methyl-D-aspartate receptors (NMDAR) play an important role in development and are also known to be associated with certain brain disorders. Commercially available NMDAR subunit specific antibodies from Cell Signaling and Abcam were shown to not be very specific and can skew results. The purpose of this investigation was to create a green fluorescent protein (GFP) tagged NR2B expressing vector that could be used to more accurately measure NMDAR2B trafficking. Following the successful creation of the GFP tagged NMDAR2B vector, the transfection protocol for this specific vector was optimized. The growth rate of the transfected cells and NMDAR2B protein expression of the transfected cells were measured in order to determine the most efficient protocol. Preliminary results of the transfection protocol suggest that changes in concentrations of chemicals used in the protocol can increase protein expression; these are being confirmed. Creating a vector with NMDAR2B and GFP will allow for examination of NMDAR2B expression in live cells, both in vitro and in vivo, and will provide a greater understanding of NMDAR function.
Creating an N-Methyl-D-Aspartate Receptor 2B-Specific Vector with Green Fluorescent Protein and Optimizing a Transfection Protocol
B-131 Grainger
N-methyl-D-aspartate receptors (NMDAR) play an important role in development and are also known to be associated with certain brain disorders. Commercially available NMDAR subunit specific antibodies from Cell Signaling and Abcam were shown to not be very specific and can skew results. The purpose of this investigation was to create a green fluorescent protein (GFP) tagged NR2B expressing vector that could be used to more accurately measure NMDAR2B trafficking. Following the successful creation of the GFP tagged NMDAR2B vector, the transfection protocol for this specific vector was optimized. The growth rate of the transfected cells and NMDAR2B protein expression of the transfected cells were measured in order to determine the most efficient protocol. Preliminary results of the transfection protocol suggest that changes in concentrations of chemicals used in the protocol can increase protein expression; these are being confirmed. Creating a vector with NMDAR2B and GFP will allow for examination of NMDAR2B expression in live cells, both in vitro and in vivo, and will provide a greater understanding of NMDAR function.