Thiskitisdesignedfortwopurposes:1)identificationofproteinsthatspeciallybindK48polyUbchainsinwholecellortissuelysates;2)validationofdirectinteractionsbetweenK48polyUbchainsandtheirbindingproteins.
TheNon-cleavableK48polyUbchain(2-4)areresistantofdeubiquitinationwhenincubatingwithwholecellortissuelysates,thusallowingmaximallycaptureproteinsthatspecificallybindK48polyUbchains.Afterbinding,6xHis-Non-cleavableK48polyUbchainsandthebindingproteinscanbeenrichedbyNickelXPureAgaroseResin(included).Theboundproteinsandthenon-cleavableK48polyUbchainscanbeelutedfromNiresinusingabuffercontaining250mMimidazole(included)orbyincubatingwiththrombin(notincluded).Eitheroftheelutionmethodcansignificantlyreducetheamountsofnon-specificbindingproteinswhencomparedwithelutionusingSDSsamplebuffer.
AdditionalInformation
| ProductName: | K48polyUbChain-bindingProteinIdentification/ValidationKit |
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| AlsoKnownAs: | N/A |
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| CatalogNo.: | J2500 |
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| Size: | 1Kit |
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| MolecularWeight: | N/A |
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| Species: | Human |
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| Source: | Bacterialrecombinant |
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| Stock: | 20mMTris,150mMNaCl,2mMβME,10%Glycerol |
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| Concentration: | Seetubelabel |
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| QualityAssurance: | 95%basedonSDS-PAGE |
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| Storage: | Seethestorageconditiononthelabelofeachcomponent;avoidmultiplefreeze-thawcycles |
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| PDFDataSheet: | PDFdatasheet |
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| NCBIRefSeq: | N/A |
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| Image(s): | No |
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| ShippingMethod: | Dryiceshipping |
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| References: | N/A |
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Details
*Thiskitisdesignedfortwopurposes:1)identificationofproteinsthatspeciallybindK48polyUbchainsinwholecellortissuelysates;2)validationofdirectinteractionsbetweenK48polyUbchainsandtheirbindingproteins.
Description
TheNon-cleavableK48polyUbchain(2-4)areresistantofdeubiquitinationwhenincubatingwithwholecellortissuelysates,thusallowingmaximallycaptureproteinsthatspecificallybindK48polyUbchains.Afterbinding,6xHis-Non-cleavableK48polyUbchainsandthebindingproteinscanbeenrichedbyNickelXPureAgaroseResin(included).Theboundproteinsandthenon-cleavableK48polyUbchainscanbeelutedfromNiresinusingabuffercontaining250mMimidazole(included)orbyincubatingwiththrombin(notincluded).Eitheroftheelutionmethodcansignificantlyreducetheamountsofnon-specificbindingproteinswhencomparedwithelutionusingSDSsamplebuffer.
Components
•6xHis-Non-cleavableK48polyUbchain(2-4)[Storeat-80°C]..........100µg
•6xHis-Ubiquitin[Storeat-80°C].............................................................................100µg
•NickelXPureAgaroseResin(sUSPendedin50%slurry)[Storeat2-8°C]....................500µl
•4MImidazole[Storeat-80°C]......................................................................................500µl
•Iodoacetamide(IAA)[Storeat-80°C]..........................................................................25mg
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