I have transfected HEK293 cells with NR1/2 subunits. I am currently using 1mM of NMDA in the activation buffer. Is this value too high/too low?
Thanks
Take a look at the attached method on patch clamping and stimulating NMDA receptors in HEK293 cells. It seems you need a higher dose than 1 mM.
http://scigine.com/method.php?ID=374598
Thank you. I think the protocol you sent might not be suitable as it describes patch clamping. I activate my NMDARs on 6 well plates, I am guessing that differs?
If you want fast and immediate receptor activation, I believe you'll need a high dose like the patch clamp method. Perhaps you'll need to optimize based on the rest of your assay. What are you using the cells for ?
I use the cells for further analysis using Western Blotting looking at total NMDAr expression after activation vs non- activated cells on 6 well plate.
First, you probably should do the calcium imaging to monitor the level of NMDA receptor activation. And it is also not hard to do a proper dose response curve in HEK cells transfected with NMDA receptors :-). If you are looking for changes in receptor internalization, a dose response curve probably is more critical,
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