Hello everyone, I am a lab technician with no flow cytometry experience whatsoever. I am hoping to develop a simple bioassay to quantitate glutamate toxicity in differentiated P19 cells. Differentiated P19 cells express functional NMDA receptors. The ligand for this receptor is glutamate. however at high levels glutamate acts as a neurotoxin and the resulting influx of calcium through the receptor channel is fatal to the cell. The assay I am hoping to develop will quantitatively examine the protective effects of selective NMDA receptor channel blockers. I plan to use facscan analysis to determine the percentage of dead cells within populations of cells that have or have not been treated with the blockers prior to addition of a fatal dose of glutamate. I could use TUNEL staining or some other marker of cell death but this is quite a long-winded affair due to the involvement of several antibody incubation steps. Is there a quicker alternative. Thanks for taking the time to read this. I hope some of you have some ideas, Wymond Symes ---------------------- Charles Wymond Symes CNS Gene Therapy Centre Dept. Molecular Medicine University of Auckland Private Bag 92019 Auckland ph 3737599 xtn 4482 fax 3737492
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