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苏州工业园区生物纳米园A4#216
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4000-520-616 / 18915418616
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0512-67156496
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商品描述
GlutathioneS-transferase(GST)genefusionsystemshavebeenwidelyusedforobtaininglargeamountsofdesirableproteininEscherichiacoli.
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| Description: GlutathioneS-transferase(GST)genefusionsystemshavebeenwidelyusedforobtaininglargeamountsofdesirableproteininEscherichiacoli.Thefusionprotein,whichcontainsaGSTtailcanthenbepurifiedthroughaffinitychromatography.MCLABsglutathioneagaroseisdesignedforthespecificpurificationofGSTrecombinantproteinsandotherglutathione-bindingproteins.Glutathioneagaroseisuniquelyformulatedforexcellentbindingcapacityandpurityoftheproteinofinterest. Protocol: ThefollowinginstructionsforGST-fusionproteinpurificationcanbescaledupordowndependingontheuserspreference.Thismanualexemplifiessamplepreparationfromaspecificamountofstartingmaterialandpurificationusing1mlresin. 1.Centrifugesampleaftercelllysistoremoveundissolvedmembranesandcellulardebrisbeforeapplyingtopurificationcolumn. 2.Washpurificationcolumnwith10xbeadvolumeofBindingBuffertoremoveazide. 3.Diluteanappropriateamountsamplewitha1:1ratioofBindingBufferbeforeapplyingtopurificationcolumn. 4.Washthepurificationcolumnwith10xbeadvolumeofBindingBufferoruntilnoproteinscanbedetectedinthewashes. 5.Eluteboundproteinofinterestwith5xbeadvolumeofElutionBuffer. 6.GSTagarosebeadscanbesavedforlaterusebywashingthepurificationcolumnwithBindingBuffercontaining3MNaCl.Afterathoroughwash,thepurificationcolumnshouldbeequilibratedinBindingBuffercontaining2mMsodiumazideandstoredat4°C. GST-fusionproteinsaregenerallyelutedfromglutathioneagarosebeadswiththeuseofexcessglutathione.Alternatively,GST-fusionproteincanbeencodedwithacleavagesitebetweentheGSTandtheprotein,allowingthedesirableproteintobeelutedwiththeuseofaprotease. RecommendedStorageConditions:-20ºC. BindingBuffer: 50mMTris 150mMNaCl pH8.0 ElutionBuffer: 50mMTrispH7.8 0.15MNaCl 1mMEDTA 1mMDTT 10mMGST |


