ab83389 Glutamate Assay Kit - Abcam

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5. Data AnalysisCorrect background by subtracting the value derived from thebackground control from all sample readings (The backgroundreading can be significant and must be subtracted from samplereadings). Plot Glutamate standard curve. Glutamate concentrationsof the test samples can then be calculated:Concentration = Sa / Sv nmol/μl, or mMWhere:Sa is the sample amount of unknown (in nmol) from standard curve,Sv is sample volume (μl) added into the wells.L-Glutamic acid Molecular Weight is 147.13 g/mol.9
6. TroubleshootingProblem Reason SolutionAssay notworkingUnexpectedresultsAssay buffer atwrong temperatureProtocol step missedPlate read atincorrect wavelengthUnsuitable microtiterplate for assayMeasured at wrongwavelengthSamples containimpeding substancesUnsuitable sampletypeSample readings areoutside linear rangeAssay buffer must not be chilled- needs to be at RTRe-read and follow the protocolexactlyEnsure you are usingappropriate reader and filtersettings (refer to datasheet)Fluorescence: Black plates(clear bottoms);Luminescence: White plates;Colorimetry: Clear plates.If critical, datasheet will indicatewhether to use flat- or U-shapedwellsUse appropriate reader and filtersettings described in datasheetTroubleshoot and also considerdeproteinizing samplesUse recommended samplestypes as listed on the datasheetConcentrate/ dilute samples tobe in linear range10
Page 7 and 8: B. Additional Materials RequiredMic
Page 9: The Glutamate Enzyme Mix is stable
Page 13: Standardcurve is notlinearNot fully

ab83389 Glutamate Assay Kit - Abcam

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