For ResearchUseOnly. NotForUseInDiagnostic Procedures. © 2013 Cell Signaling Technology, Inc. SignalSilence®, CST™, and Cell Signaling Technology® are trademarks of Cell Signaling Technology, Inc. Store at -20°C Description: SignalSilence Species Cross-Reactivity:H ecules intothecell.AllSignalSilence ofdoublestrandedRNAmol- silenced throughthedelivery ence, amethodwherebygeneexpressioncanbeselectively specifically inhibitGRB2expressionusingRNAinterfer Signaling Technology (CST)allowstheresearcherto 100 nMSignalSilence Directions forUse:CSTrecommendstransfectionwith regulates downstreamsignaling(2). and EGFRabolishesitsassociationwithSosnegatively ofTyr209pathways. Phosphorylation ofGRB2byBcr-Abl to theactivationofMAPkinaseandothersignaling which stimulatestheGTPbindingactivityofRas,leading Gab1 (1).TheSH3domainofGRB2associateswithSos, proteinssuchasEGFR,IRS-1,Shcand phosphorylated transduction. TheSH2domainofGRB2bindstotyrosine (GRB2) isaadaptorproteinthatinvolvedinRTK signal Background: Growthfactorreceptor-binding protein2 reduce targetproteinexpressionbywesternanalysis. CST arerigorouslytestedin-houseandhavebeenshownto Dg—dog Species Cross-Reactivity Key: lot-to-lot consistency. toensuremaximum previous lotbymassspectrometry composition oftheduplex.Eachlotiscomparedto toverifytheexact is furtheranalyzedbymassspectrometry affinity-solid phaseextraction.TheannealedRNAduplex coupling efficiency. Theoligoissubsequentlypurified by base bythroughtritylanalysistoensureappropriate Quality Control:Oligonucleotidesynthesisismonitored per well. transfection ina24-wellplatewithtotalvolumeof300 corresponds toafinalsiRNAconcentrationof100nMper Each vialcontainstheequivalentof100transfections,which feel freetocontactCSTwithanyquestionsonuse. provided bythetransfectionreagentmanufacturer. Please to celllysis.Fortransfectionprocedure,followprotocol Applications Key: #13265 SignalSilence Pg—pig Sc—S. cerevisiae W—Western ® GRB2siRNAI48to72hoursprior ® H—human GRB2 siRNAIfromCell IP—Immunoprecipitation Ce—C. elegans ® siRNAproductsfrom M—mouse ® GRB2siRNAI Hr—Horse R—rat - IHC—Immunohistochemistry μl Hm—hamster All—all species expected expression, whilethe (lower). TheGRB2Antibodyconfirmssilencingof loading control. with 100nMSignalSilence Western blotanalysisofextractsfrom293Tcells,transfected #6568 (-)orSignalSilence Antibody #3972(upper)or Mk—monkey 100 140 200 kDa 40 50 20 30 40 50 60 80 ChIP—Chromatin Immunoprecipitation b -Actin (D6A8)RabbitmAbisusedasa Species enclosedinparentheses are predictedto reactbasedon 100%homology. – ® ® b GRB2siRNAI(+),using ControlsiRNA(Unconjugated) Mi—mink -Actin (D6A8)RabbitmAb#8457 rev. 10/17/13 + GRB2 siRNA β-Actin GRB2 C—chicken Dm—D. melanogaster IF—Immunofluorescence and store at -20ºC. at store and Background References: of recommendedcompanionproducts. Please visitwww.cellsignal.com foracompletelisting Swiss-Prot Acc.#P62993 Entrez-Gene ID#2885 Storage: GRB2siRNAIissuppliedinRNAse-freewater. (2)  (1)  Li, S.etal.(2001)EMBOJ20,6793-804. Pawson, T. andScott,J.D.(1997)Science278,2075-80. X—Xenopus Support Orders Web F—Flow cytometry Z—zebrafish n n n www.cellsignal.com [email protected] 877-678-TECH (8324) [email protected] 877-616-CELL (2355) E-P—ELISA-Peptide B—bovine Aliquot Aliquot