The Glutamate-GloTM assay kit (Promega, J7021) was used to measure intracellular glutamate levels. Briefly, the cells were seeded in a 96-well plate, followed by glucose starvation as described above. Cells were then washed with ice-cold PBS and lysed with 0.6 n HCl. The lysing reagent wa...
After 24 h of MK-801 treatment, cells were lysed and luciferase activity was measured using the Dual-Glo Luciferase Assay System (Promega) with Luminoskan (Labosystems, Tokyo Japan). Variations in transfection efficiency between samples were normalized against the luminescence of a control reporter....
After 24 h of MK-801 treatment, cells were lysed and luciferase activity was measured using the Dual-Glo Luciferase Assay System (Promega) with Luminoskan (Labosystems, Tokyo Japan). Variations in transfection efficiency between samples were normalized against the luminescence of a control reporter....
The DRAK2 kinase reaction was per- formed in a final assay volume of 3.4 μL using the ADP-GLO kinase assay kit (Promega, Madison, WI, USA), according the ADP-GLO protocol and was read on an EnVision plate reader (Perkin Elmer, Wellesley, MA, USA). The recombinant PKCε, IKKβ, ...
The amount of GTP remaining after 15 minutes incubation at 20°C was finally assessed using the GTPase-Glo assay (Promega). The signal was normalized in each case to that in the absence of the receptor (100%). Cell culture, transfections, and inositol phosphate one (IP1) accumulation ...
HRP was visualized using LumiGLO Reagent (Cell Signaling, Danvers, MA) and Ultra Western Lightning Ultra (Perkin-Elmer) and images were captured using a Luminescence Image Analyzer (LAS-4000; FujiFilm Life Science, Stamford, CT) and analyzed using Image Quant software (GE Healthcare, Baulkham ...
Glutamine/Glutamate-Glo™ Assay Part Numbers:J8021,J8022 Sensitive, Quick Detection of Glutamine and Glutamate in Biological Samples Detect even small changes in metabolites Multiplex with other metabolite or viability assays Perform on a variety of sample types ...
Intracellular NADPH level and GSH/GSSG ratio were determined using fluorimetric SensoLyte NADP/NADPH Assay (AnaSpec) and GSH/GSSG-Glo (Promega), respectively. Intracellular Metabolite Measurements The intracellular levels of α-KG, fumarate, succinate, and malate were determined by using commercial ...
(2014) [56] found that LY341495 was behaving as inverse agonist after pre-treatment with agonist, when using the Glo-sensor cAMP assay with CHO-K1 cells stably expressing the mGlu2 receptor. Moreover, inverse agonism by LY341495 at the mGlu2 receptor was also seen in studies investigating ...
Total cellular glutathione was measured using the GSH/GSSG-Glo Assay Kit. Briefly, cells were seeded at 1000 cells/well in 96 well plates after 48 hr of siRNA transfection and treated with DMSO or Torin1 for an additional 24 hr. After treatment media was removed and cells were lysed on a...