no, it isn't. it would not stay easily in the gel and it won't bind to any membrane. for single aminoacids, chromatografic methods are used. but for quantifications, it is better to go to liquid chromatorgraphy; elisa may work, but will be very noisy.
no, it isn't. it would not stay easily in the gel and it won't bind to any membrane. for single aminoacids, chromatografic methods are used. but for quantifications, it is better to go to liquid chromatorgraphy; elisa may work, but will be very noisy.
Hi! If you are looking for a reliable and cost-effective method for glutamate determination in liquids, I could recommend the Amplex Red Glutamate Kit (available from Molecular Probes but the individual components can be bought from Sigma or other sources). It is remarkably sensitive, robust and easy to handle. All you need is a microplate (or cuvette) reader capable to read fluorescent output (preferable) or colorimetry. For Glutamate in tissue (brain) I have also used an antibody (Chemicon), but quantitative conclusions are difficult. ELISAs are available but -to my knowledge- are more expensive and less sensitive than the abovementioned Amplex Red Kit.
In lysates the Amplex Red should work. Due to its sensitivity a high dilution factor of the tissue lysate is possible. Centrifugation and/or filtration steps may be necessary to clear the lysate from interfering debris. Please note, that in tissue lysates you will detect intra-AND extracellular glutamate. I assume that you are more interested in extracellular glutamate. To determine this, you would have had to perform microdialysis. I usually measure (extracellular) glutamate in cell culture supernatant. You should check, wether this combined measure of EC and IC glutamate makes any sense.
well, first you should ask why you want to measure glutamate in brain homogenates. If you want to measure neurotransmitter glutamate, your measure is basically meaningless. If you want to measure metabolic glutamate, you should pay attention to tissue preparation and standard homogeneates may be not good.
I was reading the thread and impressed by the inputs. If any one still following this thread, could you please tell me how could one measure the level of extracellular Glutamate released in Hippocampal cells? Thanks