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ubpbio/Rapid 26S Proteasome Purification Kit-S/J4310/1 Kit

价格
¥5600.00
货号:J4310
浏览量:127
品牌:UBPBio
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商品描述
TheRapid26SProteasomePurificationKitisdesignedtorapidlypurifytheendogenous26Sproteasomefromcellortissueextractswithin8hoursaccordingtoapublishedmethod(reference1)withmodifications.ThisapproachusestheN-terminalubiquitin-like(Ubl)domainofhumanRAD23Basanaffinitybait,whichallowstherapidandgentleisolationofendogenous26Sproteasomes.Thebound26SproteasomeissubsequentlyelutedusingtheC-terminaltwoubiquitin-interactingmotifs(UIM)ofhumanS5a.

AdditionalInformation

Details

*Thiskitissufficientforpurificationofthe26Sproteasomefrom50mgor5X10mgcellortissueextractsataconcentrationof5mg/ml.Usually,1-2%oftotalcellularproteinsarethe26Sproteasome.
Description
TheRapid26SProteasomePurificationKitisdesignedtorapidlypurifytheendogenous26Sproteasomefromcellortissueextractswithin8hoursaccordingtoapublishedmethod(reference1)withmodifications.ThisapproachusestheN-terminalubiquitin-like(Ubl)domainofhumanRAD23Basanaffinitybait,whichallowstherapidandgentleisolationofendogenous26Sproteasomes.Thebound26SproteasomeissubsequentlyelutedusingtheC-terminaltwoubiquitin-interactingmotifs(UIM)ofhumanS5a.
Components
•GST-Ubl(4mg/ml)...................................................0.6ml
•GST(4mg/ml)..........................................................0.5ml•6xHis-UIM(4mg/ml)..............................................0.2ml
•20XLysisBuffer........................................................1.2ml•NaCl(1M)...............................................................1.25ml
•ATP(500mM)............................................................0.1ml•SUC-LLVY-AMC(50mM)..........................................10µl
•GlutathioneAgaroseResin(75%slurry)...........1.0ml•NickelXPureAgaroseResin(50%slurry)..........500µl
Images(Clickimagetoenlarge)

A.CoomassiestainedSDS-PAGEof5μgpurifiedGST-Ubl.B.CoomassiestainedSDS-PAGEof5μgpurified6xHis-UIM.
C.CoomassiestainedSDS-PAGEof3μgpurifiedhuman26Sproteasomes.D.Coomassiestainednative-PAGEof3μgpurifiedhuman26Sproteasomes.
E.Solutionpeptidaseassayofthepurifiedhuman26Sproteasome.150μlreactionmixturescontained10nMpurifiedhuman26Sproteasomein50μMSUC-LLVY-AMCin20mMTris,pH7.1at37°C,2mMμME.ThereleasedAMCfluorescencewasmeasuredusingaplatereaderwithexcitationandemissionfiltersat360/40and460/30nm,respectively.Thisassaywasusedtomonitorthechymotrypsin-likeactivityofthe26Sproteasome.(Clickimagetoenlarge)
ProductName:Rapid26SProteasomePurificationKit-S
AlsoKnownAs:N/A
CatalogNo.:J4310
Size:1Kit
MolecularWeight:N/A
Species:N/A
Source:N/A
Stock:Enzymestockedin20mMTris,150mMNaCl,2mMβME,10%Glycerol;SubstratestockedinDMSO
Concentration:Seetubelabel
QualityAssurance:Enzyme>95%bySDS-PAGE;Substrate>98%byHPLCandNMR
Storage:Storeat-80°C;avoidmultiplefreeze-thawcycles
PDFDataSheet:PDFdatasheet,MSDS
NCBIRefSeq:N/A
Image(s):(Clickimagetoenlarge)
Schematicof26Sproteasomeenrichment
ShippingMethod:Dryiceshipping
References:1.BescheHC,HaasW,GygiSP,GoldbergAL.Biochemistry.2009;48:2538-49.
2.ElsasserS,SchmidtM,FinleyD.MethodsEnzymol.2005;398:353-63.
3.KisselevAF,GoldbergAL.MethodsEnzymol.2005;398:364-78.
Schematicof26Sproteasomeenrichment
UBPBio are formed by conjugating the C-terminal glycine residue of Ub onto any of seven internal lysine residues or the amino group of the previous Ub. Ub chains are classified by the lysine residue used to link Ubs; different Ub chain topologies can result in different signals. For instance, Ub chains linked through lysine 6, 11, 27, 29, 33 and 48 are capABLe of targeting proteins for proteasomal degradation; in contrast, Ub chains linked through lysine 63 or the N-terminal amino group (linear Ub chains) often play important nonproteolytic functions including regulation of kinase activation and protein translation. All Ub chain products are produced by using of human wild type Ub reacting with specific E2s.Additional InformationProduct Name:K11-Ub(n≥3)Also Known As:K11-Ub(n≥3)Catalog No.:D3301Size:50 µgMolecular Weight:N/ASpecies:HumanSource:Bacterial recombinantStock:20 mM Tris, 150 mM NaCl, 2 mM βME, 10% GlycerolConcentration:See tube labelQuality Assurance:~95% by SDS-PAGE, see datasheet for gel imageStorage:Store at -80°C; avoid multiple freeze-thaw cyclesPDF Data Sheet:PDF Datasheet, MSDSNCBI RefSeq:N/AImage(s):N/AShipping Method:Dry ice shippingReferences:1. Hershko A, et al. (1980)Proc Natl Acad Sci USA 77(4), 1783 – 1786.2. Pickart CM, (1997) FASEB 11(13), 1055 – 1066.3. Hershko A, et al. (1998) Ann Rev Biochem 67, 425 – 479.4. Jiang X, et al. (2012) Nature Reviews Immunology 12, 35 – 48.DetailsUb chains are formed by conjugating the C-terminal glycine residue of Ub onto any of seven internal lysine residues or the amino group of the previous Ub. Ub chains are classified by the lysine residue used to link Ubs; different Ub chain topologies can result in different signals. For instance, Ub chains linked through lysine 6, 11, 27, 29, 33 and 48 are capable of targeting proteins for proteasomal degradation; in contrast, Ub chains linked through lysine 63 or the N-terminal amino group (linear Ub chains) often play important nonproteolytic functions including regulation of kinase activation and protein translation. All Ub chain products are produced by using of human wild type Ub reacting with specific E2s.