Cov khoom

Tris acetate ntsev feem ntau siv rau hauv kev npaj ntawm TAE (Tris-acetate-EDTA) tsis, uas yog siv los ua kev khiav tsis zoo thiab hauv agarose gels.Tris Acetate-EDTA tsis yog siv rau DNA agarose gel electrophoresis tab sis kuj yog siv rau cov tsis-denaturing RNA agarose gel electrophoresis.Ob-stranded DNA zoo li khiav nrawm dua hauv TAE dua li lwm cov buffers thiab tej zaum yuav qaug zog thaum txuas ntxiv electrophoresis.Tsis siv neeg los yog hloov tsis tau thaum lub sij hawm txuas ntxiv electrophoresis tuaj yeem kho qhov muaj peev xwm qis dua.Tej zaum yuav raug siv ntawm ntau qhov ntau los kawm txog kev txav mus los ntawm DNA hauv kev daws teeb meem.Txij li thaum borate nyob rau hauv TBE tsis (Tris-Borate-EDTA Buffer, 10X Powder Pack, sc-296651) yog ib tug muaj zog inhibitor rau ntau enzymes, TAE tsis yog pom zoo thaum saib enzymatic daim ntaub ntawv rau cov qauv DNA.Tris acetate ntsev kuj yog ib qho tsis muaj qhov siab tshaj plaws hauv ATP kev soj ntsuam nrog firefly luciferase.

Siv: TAE khiav tsis yog feem ntau siv tsis rau DNA agarose gel electrophoresis, thiab tseem siv rau haiv neeg RNA agarose gel electrophoresis.Ob-stranded DNA nyiam txav nrawm dua hauv TAE dua li lwm cov buffers, tab sis kuj ua luam dej tsis zoo vim qhov tsis txaus ntawm cov ions tsis zoo thaum lub sij hawm ntev electrophoresis.Tsis siv neeg caij tsheb kauj vab lossis tsis sib pauv thaum lub sij hawm ntev electrophoresis tuaj yeem them rau qhov tsis muaj peev xwm qis dua.2 Dilute cov concentrated TAE tsis tau kom tau 1 mMTAE tsis muaj 40 mM Tris acetate thiab 1 mM EDTA, pH 8.3.1 mMTAE tsis tuaj yeem siv tau ob qho tib si hauv agarose gels thiab ua haujlwm tsis ua haujlwm.Rau qhov kev daws teeb meem siab tshaj plaws, nws raug pom zoo kom siv qhov hluav taws xob qis dua 5V / cm (qhov nrug ntawm chav tsev electrodes).

Daim ntawv thov: TAE khiav tsis yog feem ntau siv tsis rau DNA agarose gel electrophoresis nyob rau hauv Chemicalbook gel, thiab nws kuj yog siv rau non-denaturing RNA agarose gel electrophoresis.Ob-stranded DNA nyiam txav nrawm dua hauv TAE dua li lwm cov buffers, tab sis kuj ua luam dej tsis zoo vim qhov tsis txaus ntawm cov ions tsis zoo thaum lub sij hawm ntev electrophoresis.Tsis siv neeg caij tsheb kauj vab lossis tsis sib pauv thaum lub sij hawm ntev electrophoresis tuaj yeem them rau qhov tsis muaj peev xwm qis dua.Cov concentrated TAE tsis yog diluted kom tau 1 mMTAE tsis muaj 40 mM Tris acetate thiab 1 mM EDTA, pH 8.3.1 mMTAE tsis tuaj yeem siv tau ob qho tib si hauv agarose gels thiab ua haujlwm tsis ua haujlwm.Rau qhov kev daws teeb meem siab tshaj plaws, nws raug pom zoo kom siv qhov hluav taws xob qis dua 5V / cm (qhov nrug ntawm chav tsev electrodes).

Siv: Hauv kev tshawb pom ntawm ATP nrog firefly luciferase, cov khoom no yog qhov tsis zoo tshaj plaws;glutamate khi nrhiav kom tau.

Kev sib cais ntawm [3H] l-glutamic acid rau cov ntaub ntawv tsis-receptor.

[3H] L-glutamic acid khi rau microfuge hlab thiab iav tau tshawb xyuas hauv plaub qhov tsis sib xws.Cov ntaub ntawv keeb kwm yav dhau los khi rau cov ntaub ntawv no tsis tsim nyog, tab sis tau nce los ntawm centrifugation lossis suction hauv Tris-HCl thiab Tris-citrate buffer.Qhov kev khi no tsawg dua lossis Thaum HEPES-KOH, lossis Tris-acetate tsis siv.[3H] L-glutamate khi rau microfuge hlab yog inhibited los ntawm L- tab sis tsis yog D-isomers ntawm glutamate thiab aspartate.DL-2-amino-7-phosphonoheptanoic acid kuj tsis cuam tshuam kev khi.Lwm qhov sib xyaw ua ke uas pom tias muaj kev cuam tshuam qis rau nruab nrab yog: N-methyl-D-aspartate, quisqualate, L-glutamic acid diethyl ester, N-methyl-L-aspartate, kainate, thiab 2-amino-4-phosphonobutyrate.Kev khi tau inhibited los ntawm denatured nas hlwb membranes.Ib qho protein-dependent [3H] glutamate binding tau txais nrog kev rov ua kom khov-thawed membrane ntau zaus thaum khi tau ua tiav hauv Tris-acetate tsis.Nws raug pom zoo tias Tris-acetate lossis HEPES -KOH tsis yuav tsum tau siv rau hauv glutamate bin ding assay.Yog tias Tris-HCl lossis Tris-citrate tsis yog siv, kev sim tswj kom tsim nyog yuav tsum tau ua kom raug kho rau kev khi rau microfuge hlab lossis iav fiber ntau lim.