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.
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.
SouthernBiotech/Goat Anti-Human Lambda-Alexa Fluor® 555/2070-32/1.0 mg
vectorlabs/Biotinylated Aleuria Aurantia Lectin (AAL)/B-1395/1 mg
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Mouse IgG)/PK-6102/1 kit
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Universal)/PK-6200/1 kit
vectorlabs/Unconjugated Musa Paradisiaca (Banana) Lectin (BanLec)/L-1410/5 mg
GiottoBiotech/Calmodulin N60D/5 mg/G02CLM60cn