Assay Method Information |
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| Microplate GCPII Radioactivity-Based Assay |
Description: | GCPII activity in vitro is monitored through the hydrolysis [3H]NAAG to NAA and [3H]Glu. The radioactivity-based assay was miniaturized to a 96-well format. After the reaction, an aliquot of the reaction mixture was transferred to a 96-well spin column containing AG1X8 ion-exchange resin; the plate was then centrifuged. NAA[3H]G bound to the resin and [3H]G eluted in the flowthrough. The flowthrough and the washes were collected, and was transferred to a solid scintillator-coated 96-well plate (Packard) and dried to completion. The radioactivity corresponding to [3H]G was determined with a scintillation counter. Assay points for each experiment were mostly the average of eight determinations. |
Affinity data for this assay | |
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