BradfordAssay
...ample#5(0.8mg/mL):24uLBSA+6uLbuffer.Sample#6(1.0mg/mL):30uLBSA+0uLbuffeer.*Note:TheubiquitouslyusedBSAisaverypoorstandardforthebradfordassay.Toadjustforthis,multiplytheresultsyougetforyourproteinconcentrationby2.1togetacloserapproximationofyourprotein‘sconcentration.Lysozyme,ovalbuminandcatalasema...
藥品天地;專業藥學;實驗技術;基本實驗技術;生物分子定量BradfordProteinConcentrationAssay
...tandardsandtheproteinsampletobetestedaccordingtoTable1indisposablecuvettes.Notethatthereferencecellandblankareidentical.Areferencecelltestsampleisonlyrequiredwhenusingadouble-beamUV-visiblespectrophotometerforabsorbancemeasurements.Notethatadilutionoftheproteinsamplemayberequiredfortheresultingabsor...
藥品天地;專業藥學;實驗技術;基本實驗技術;生物分子定量UseoftheBradfordProteinAssayinaMicrotiterPlateFormat
...tandardshouldbedissolvedinabuffersimilartothattheunknownswillbedissolvedin.Note:otherstandardsareacceptableandmaybepreferredforcertainapplications(forexample,useanIgGstandardwhenmeasuringconcentrationsofimmunoglobulins.Microtiterplates-weuseMicroTestIIIFlexibleassayplatefromFalconbecausetheyarecheap...
藥品天地;專業藥學;實驗技術;基本實驗技術;生物分子定量UVAbsorbance(280nm)–ProteinDetermination
...idwaytocalculatetheextinctioncoefficientisusingtheProtPARAMtoolfromExPASy.(Note:youneedtoknowtheprotein‘saminoacidsequenceinordertocalculatetheextinctioncoefficient.)InterferingReagentsforUVAbsorbance280nmREAGENTCONCENTRATIONAmmoniumSulfate50%Brij351%DTT3mMEDTA30mMGlycerol40%KCl100mMßME10mMN...
藥品天地;專業藥學;實驗技術;基本實驗技術;生物分子定量SpotFluorescence法定量DNA
...lebycomparingtheintensityofthefluorescenceinthesampleswiththestandarddrops.NOTE:TheDNAstandardsshouldbesimilarinstructuretothesampleyouwishtomeasure.Workveryfasttoavoidanyevaporationfromthedropsbeforethephotographismade.下載全文作者:佚名
藥品天地;專業藥學;實驗技術;基本實驗技術;生物分子定量AngiotensinProteinKinaseAssay
...ndcount.Val5-angiotensinIIisobtainedfromSigmaandstoredasa50mMstockat-70°C.Note:Ifyougetmorethan100,000cpmincorporated,youmaybepeggingtheassay.Inthiscase,repeattheexperimentanduselessoftheprecipitateintheassay.ConsumptionofmostoftheATPcanoccurifyouarestudyingproteinsover-expressedbytransienttransfec...
藥品天地;專業藥學;實驗技術;生化實驗;氨基酸和蛋白實驗Glycosphingolipidanalysis
...lofchloroformtobringthesolventratioto3:6:0.8ofchloroform-methanol-water.---NOTE:Itisbestnottoextractatthispointbecausegangliosidescouldpotentiallypartitionintotheaqueousphaseandbelost.---Thismixturecanbestoredinthefreezerforshortperiodsoftime.9)PrepareaslurryofDEAE-Sephadexinchloroform-methanol-wate...
藥品天地;專業藥學;實驗技術;生化實驗;脂肪實驗Lipidanalysis:EXTRACTIONOFLIPIDSFROMLIPOPHORIN
...:2:1.8,nowthepolarandnonpolarsolventsarenolongermiscible,andwegettwophases.Note:Chloroformisheavierthanwater.Thespeedandeaseofthismethodmakeitsuitableformultipleextractions,e.g.ofplasmasamples,providedthatasmallamountofofaqueouscontaminationcanbetolerated.Experimentalprotocol:Useonlyglasspipettest...
藥品天地;專業藥學;實驗技術;生化實驗;脂肪實驗MethylationofFattyAcids(KropinskiMethod)
...-HClreagent/internalstandardintoeachtubeandvortex.Heatat100oCfor20minutes.(Note:eachtubeshouldbeverywellsealedwithteflontapeandgreasetopreventevaporation.)Sonicateandheatat100oCovernight.Neutralizeaciditywith0.5NNaOH.TestpHwithpHpaper.Centrifugeinclinicalcentrifugefor5minutes.Savesupernatentincleang...
藥品天地;專業藥學;實驗技術;生化實驗;脂肪實驗NeutralizingArachidonicAcid
NeutralizingArachidonicAcid:Contributor:SupryaJayadevDate:August21,1991NOTE-UsearachidonicacidfromBiomol(#FA-003)1)MakeuptheappropriateconcentrationofAAin100%ethanol.2)Add1µlofphenolredsolution.--Thesolutionshouldappearyellowsincethefreeacidhasnotbeenneutralizedasyet.3)Add1µlatatimeof5...
藥品天地;專業藥學;實驗技術;生化實驗;脂肪實驗