For ResearchUseOnly. NotForUseInDiagnostic Procedures. ® 2015 Cell Signaling Technology, Inc. XP and Cell Signaling Technology are trademarks of Cell Signaling Technology, Inc. Store at -20°C Dg—dog Species Cross-Reactivity Key: Applications Key: labeled withDyLight™554Phalloidin#13054 (red). HEXIM1 (D5Y5K)RabbitmAb(green).Actin filaments were Confocal immunofluorescentanalysisofHeLa cellsusing protein. ing toresiduessurroundingGly63ofhumanHEXIM1 by immunizinganimalswithasyntheticpeptidecorrespond Source/Purification: Monoclonalantibodyisproduced recognizes endogenouslevelsoftotalHEXIM1protein. Specificity/Sensitivity: HEXIM1(D5Y5K)RabbitmAb coid receptorregulatedgenes(9-12). gene expressionprogramssuchasestrogenandglucocorti play acriticalroleinregulationofwiderangecellular active P-TEFbinthecellbyHEXIM1/7SKRNAisthoughtto tion (2,3).Theregulationoftherelativeratioinactiveto P-TEFb inaninactiveformleadingtotranscriptioninhibi interacts withthecyclinT1subunitofP-TEFb,sequestering of transcriptionelongation(4-8).7SKRNA-boundHEXIM1 subunit ofRNApolymeraseIIandisanimportantregulator theC-terminaldomainoflargest P-TEFb phosphorylates of positivetranscriptionelongationfactorb(P-TEFb)(2,3). non-coding RNA,andtogethertheyactasapotentinhibitor sacetamide (1).HEXIM1binds7SKRNA,ahighlyabundant treatment withthedifferentiatingagenthexamethylenebi smooth musclecellsasaproteinthatisupregulatedupon protein 1(HEXIM1)wasoriginallyidentifiedinvascular Background: Hexamethylenebis-acetamide-inducible #12604 0.1% Tween IMPORTANT: Forwesternblots,incubatemembrane withdilutedantibodyin5%w/vBSA,1XTBS, n HEXIM1 (D5Y5K)RabbitmAb HeLa 3 Endogenous Applications W, IP, IF-IC Pg—pig  (10 westernblots) 100 µl ® Sc—S. cerevisiae 20 at4°Cwithgentle shaking,overnight. W—Western H—human Species Cross-Reactivity* IP—Immunoprecipitation Ce—C. elegans H, Mk M—mouse

Hr—horse R—rat IHC—Immunohistochemistry - - - - Hm—hamster Rabbit (DA1E)mAbIgGXP Immunoprecipitation ofHEXIM1fromHeLacellextracts,using Rabbit mAb. Western blotanalysiswasperformedusingHEXIM1(D5Y5K) HEXIM1 (D5Y5K)RabbitmAb(lane3).Lane1is10%input. HEXIM1 (D5Y5K)RabbitmAb Western blotanalysisofextractsfromvariouscelllinesusing Molecular Wt. 100 140 kDa 20 30 40 50 60 80 60 kDa 100 140 200 kDa 10 20 30 40 50 60 80 All—all species expected Mk—monkey 1 – – ChIP—Chromatin Immunoprecipitation + – 2 HeLa – + 3

® A-431 IsotypeControl#3900(lane2)or Mi—mink Rabbit IgG** rev. 01/05/15 HEXIM1 (D5Y5K)RabbitmAb XP Rabbit (DA1E)mAbIgG Hep G2 Isotype IgG lightchain HEXIM1 ® IsotypeControl COS-7 Species enclosed inparenthesesarepredicted toreactbasedon100% homology. C—chicken HEXIM1 Dm—D. melanogaster IF—Immunofluorescence UniProt ID#O94992 Entrez-Gene ID#10614 (12)  (11)  (10)  Background References: of recommendedcompanionproducts. Please visitwww.cellsignal.com foracompletelisting for thisproductatwww.cellsignal.com. For productspecificprotocolspleaseseethewebpage Immunofluorescence (IF-IC) 1:1200 Immunoprecipitation 1:100 Western blotting Recommended AntibodyDilutions: ** *Species cross-reactivityisdeterminedbywesternblot. sodium azide.Storeat–20°C.Donotaliquottheantibody. mM NaCl,100µg/mlBSA,50%glycerolandlessthan0.02% Storage: Suppliedin10mMsodiumHEPES(pH7.5),150 Tween isaregisteredtrademark ofICIAmericas,Inc. subsidiaries. DyLight isatrademarkof ThermoFisherScientificInc.andits (9)  (8)  (7)  (6)  (5)  (4)  (3)  (2)  (1)  detect thisantibody. antibodiesmustbeusedto Anti-rabbit secondary Peterlin, B.M.etal.(2012)WileyInterdiscipRevRNA3, Yamada, T. etal.(2006)MolCell21,227-37. Wada, T. etal.(1998)EMBOJ17,7395-403. Pirngruber, J.etal.(2009)CellCycle8,3636-42. Lenasi, T. andBarboric,M.(2010)RNABiol7,145-50. Buratowski, S.(2009)MolCell36,541-6. Yik, J.H.etal.(2003)MolCell12,971-82. Michels, A.A.etal.(2004)EMBOJ23,2608-19. Ouchida, R.etal.(2003)GenesCells8,95-107. Shimizu, N.etal.(2005)ProcNatlAcadSciUSA102, Ogba, N.etal.(2008)CancerRes68,7015-24. Ketchart, W. etal.(2011)Oncogene30,3563-9. 92-103. 8555-60. 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 1:1000