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absoluteantibody/Anti-DNA/RNA G-quadruplex (and related small molecules) [BG4]/100 μg/Ab00174-1.6188bio精品生物—专注于实验室精品爆款的电商平台 - 蚂蚁淘旗下精选188款生物医学科研用品
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absoluteantibody/Anti-DNA/RNA G-quadruplex (and related small molecules) [BG4]/100 μg/Ab00174-1.6

UniProtAccessionNumberofTargetProtein:n/a

Immunogen:DNA/RNAG-quadruplex.

Specificity:BindswithhighselectivityandlownanomolaraffinitytoDNAandRNAG-quadruplexstructures.

ApplicationNotes:Thisantibodyhasbeenshown(byELISA)tobindselectivelytoG-quadruplexformingDNAandRNAstructures.PleasenotethatunliketheoriginalscFvreportedbyBiffietal.,thisantibodydoesnothaveFLAG-tag.Weldonetal.(2016,PMID:27820800)usedAb00174-1.1toidentifyG-quadruplexesinlongfunctionalRNAsusinganelectrophoreticmobilityshiftassay(EMSA).BG4wasshowntohaveweakcross-reactivitywithiMotifstructures(2018,PMID:29686376).iMotifstructurescanselectivelybedetectedusingouranti-iMotifantibodyAb01462.

Antibodyfirstpublishedin:BiffiG, TannahillD, McCaffertyJ, BalasubramanianS.QuantitativevisualizationofDNAG-quadruplexstructuresinhumancells.NatChem. 2013Mar;5(3):182-6.PMID:23422559Noteonpublication:DescribesgenerationofaG-quadruplex-specificantibodyusingphagedisplayandtheinvitroselection(usingELISAandCD)ofastructure-specificantibodyprobethatbindswithhighselectivityandlownanomolaraffinitytoDNAG-quadruplexstructurescalledBG4.

Binding Curve
ELISAofBG4antibodybindingtoG4-formingRNAandDNAoligonucleotides.
Binding Curve

ComparisonofthebindingoffulllengthIgGandFabversionsoftheBG4antibodyBindingcurvesfortheassociationofBG4asafull-lengthmurineIgG1(leftpanel)andahumanFabfragment(rightpanel)withaDNAG-quadruplexoligo(fromhumanMYCgenepromoter:5’[Btn]TGAGGGTGGGTAGGGTGGGTAA),aRNAG-quadruplexoligo(frumhumanNRAS5’UTRRNA:5’[Btn]TGAGGGTGGGTAGGGTGGGTAA)andanon-G-quadruplexformingnegativecontrololiogo(mutatedMYCsequence:5′[Btn]TGAGAGTGAGTAGAGTGAGTAA)asdeterminedbyELISA.ToformtheG-quadruplex,theoligos(at5uM)wereannealedin10mMTris,100mMKCl(pH7.4)andheatedto95°Cfor10minutesbeforeslowlycoolingtoRT.TheELISAwasperformedbybindingthebiotinylatedoligos(50nM)toasteptavidinplatefor1hour.Afterwashing,theplatewasblockedandincubatedwithserialdilutionsofantibodyfor1houratRT.Theplateswerewashedandthenincubatedwithananti-mouseoranti-human,HRP-coupledsecondaryantibody.Afterwashing,TMBsubstratewasadded,thereactionstoppedwithHClandtheabsorbancemeasuredat450nmonaplatereader.Eachexperimentaldatapointwasdeterminedintriplicate.ThismethodisdescribedintheNatureChemistrypaper(doi:10.1038/NCHEM.1548,dos:10.1038/NCHEM.1805).DatacourtesyofDarcieMulhearnandDr.DavidTannahill,BalasubramanianGroup,DepartmentofChemistry,UniversityofCambridge.

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