Mdm4/HdmXTrap Agarose | Chromotek

Description
Mdm4/HdmX-TrapAgaroseisanaffinityresinforimmunoprecipitationofMdm4/HdmX-fusionproteins.
ItconsistsofMdm4/HdmXNanobodiesconjugatedtoagarosebeads.

Specificity
Species-Reactivity:human
Epitope:aa1-129

Applications
Immunoprecipitation(IP)/Co-IP
Massspectrometry
On-beadenzymeassays

Specificity
Doubleminute4protein(Mdm4/HdmX),human
NotethatbindingofMdm4/HdmX-Trapinterfereswithbindingofp53toMdm4HdmX.

CoupledNanobody/VHH
Recombinant,monoclonalanti-Mdm4/HdmXsingledomainantibody(sdAb)fragment

Beadproperties
Beadsize:~90µm(cross-linked4%agarosebeads)
Storagebuffer:20%EtOH

Elution
-SDSsamplebuffer
-0.2MglycinepH2.5
Insteadofelution,werecommendon-beadassayslikeon-beaddigestionformassspectrometryanalysis.

RRID
AB_2631374

Storageinstructions
Shippedatambienttemperature.Uponreceiptstoreat4°C.Stableforoneyear.

Forpreclearingofyoursamplewerecommendtouseourbindingcontrols(bab-20orBMAb-20).

PleasefindmoreinformationinourTroubleshootingguide

Youcaneluteyourfusionproteinofinterestwith0.2MglycinepH2.5atroomtemperature.Pipettethebeadsupanddownfor60-120secondsandrepeatthisstep.Ensuretoneutralizeyoursupernatantimmediatelyafterwardsbyadding1MTrisbasepH10.4.

Foroneimmunoprecipitationreaction,werecommendusing~10^6-10^7mammaliancells.Theyieldisalsodependentontheexpressionlevelofyourproteinofinterestandtheinteractionpartners.

Forothertypeofcellsthanmammaliancells,werecommendusing0.5-1.0mgofcellextract.

No,youcandirectlyconductanon-beaddigestionafterimmunoprecipitation.Thisprocedureallowsfasterandmoreefficientsamplepreparationandapotentialhigheryield.
Pleasefindmoreinformationhere:

Preomicskit

Protocolon-beaddigestion

No,youcandirectlyperformyourenzymaticassayonthebeadsiftheactivecenterisnotblocked.

Pleasefindmoreinformationinourapplicationnoteenzymaticactivityassay

25µLslurryaresufficientforonepull-downreactionastheaffinityofthetrapsisveryhigh.

TheMdm4/HdmX-TrapAgaroseisalsoavailableinakit,including:

Mdm4/HdmX-TrapAgaroseLysis*,washandelutionbuffers

*Thelysisbufferprovidedissuitableformammaliancells.Forothercelltypesuseanothersuitablelysisbuffer.

EasyandconvenienthandlingRapidwashingandcleanelutionofboundproteinsSimplifythepulldownofyourproteinSpinColumnsforMdm4/HdmX-TrapAgarose
Controlforunspecificbindingofproteins,DNA,etc.tobeadsPre-clearingofcelllysateBindingControlforMdm4/HdmX-TrapAgarose