HEXIM1 (D5Y5K) Rabbit mAb Store at -20°C 3 n 100 µl Orders n 877-616-CELL (2355) (10 western blots) [email protected] Support n 877-678-TECH (8324) [email protected] Web n www.cellsignal.com rev. 01/05/15 #12604 For Research Use Only. Not For Use In Diagnostic Procedures. Entrez-Gene ID #10614 UniProt ID #O94992 Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 Applications Species Cross-Reactivity* Molecular Wt. Isotype mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% W, IP, IF-IC H, Mk 60 kDa Rabbit IgG** sodium azide. Store at –20°C. Do not aliquot the antibody. Endogenous *Species cross-reactivity is determined by western blot. Background: Hexamethylene bis-acetamide-inducible ** Anti-rabbit secondary antibodies must be used to protein 1 (HEXIM1) was originally identified in vascular kDa HeLa A-431 Hep G2 COS-7 detect this antibody. smooth muscle cells as a protein that is upregulated upon 200 Recommended Antibody Dilutions: treatment with the differentiating agent hexamethylene bi- 140 Western blotting 1:1000 sacetamide (1). HEXIM1 binds 7SK RNA, a highly abundant 100 Immunoprecipitation 1:100 non-coding RNA, and together they act as a potent inhibitor 80 Immunofluorescence (IF-IC) 1:1200 of positive transcription elongation factor b (P-TEFb) (2,3). 60 HEXIM1 P-TEFb phosphorylates the C-terminal domain of the largest 50 For product specific protocols please see the web page subunit of RNA polymerase II and is an important regulator for this product at www.cellsignal.com. 40 of transcription elongation (4-8). 7SK RNA-bound HEXIM1 30 Please visit www.cellsignal.com for a complete listing interacts with the cyclin T1 subunit of P-TEFb, sequestering of recommended companion products. P-TEFb in an inactive form leading to transcription inhibi- 20 tion (2,3). The regulation of the relative ratio of inactive to Background References: active P-TEFb in the cell by HEXIM1/7SK RNA is thought to (1) Ouchida, R. et al. (2003) Genes Cells 8, 95-107. 10 play a critical role in regulation of a wide range of cellular (2) Michels, A.A. et al. (2004) EMBO J 23, 2608-19. gene expression programs such as estrogen and glucocorti- (3) Yik, J.H. et al. (2003) Mol Cell 12, 971-82. coid receptor regulated genes (9-12). Western blot analysis of extracts from various cell lines using HEXIM1 (D5Y5K) Rabbit mAb (4) Buratowski, S. (2009) Mol Cell 36, 541-6. Specificity/Sensitivity: HEXIM1 (D5Y5K) Rabbit mAb recognizes endogenous levels of total HEXIM1 protein. (5) Lenasi, T. and Barboric, M. (2010) RNA Biol 7, 145-50. Source/Purification: Monoclonal antibody is produced (6) Pirngruber, J. et al. (2009) Cell Cycle 8, 3636-42. kDa 1 2 3 by immunizing animals with a synthetic peptide correspond- (7) Wada, T. et al. (1998) EMBO J 17, 7395-403. ing to residues surrounding Gly63 of human HEXIM1 140 protein. (8) Yamada, T. et al. (2006) Mol Cell 21, 227-37. 100 80 (9) Peterlin, B.M. et al. (2012) Wiley Interdiscip Rev RNA 3, HEXIM1 92-103. HeLa 60 50 (10) Ketchart, W. et al. (2011) Oncogene 30, 3563-9. 40 (11) Ogba, N. et al. (2008) Cancer Res 68, 7015-24. 30 IgG light chain (12) Shimizu, N. et al. (2005) Proc Natl Acad Sci U S A 102, 8555-60. 20 – + – Rabbit (DA1E) mAb IgG XP® Isotype Control – – + HEXIM1 (D5Y5K) Rabbit mAb Immunoprecipitation of HEXIM1 from HeLa cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or HEXIM1 (D5Y5K) Rabbit mAb (lane 3). Lane 1 is 10% input. Confocal immunofluorescent analysis of HeLa cells using Western blot analysis was performed using HEXIM1 (D5Y5K) HEXIM1 (D5Y5K) Rabbit mAb (green). Actin filaments were Rabbit mAb. labeled with DyLight™ 554 Phalloidin #13054 (red). IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS, DyLight is a trademark of Thermo Fisher Scientific Inc. and its 0.1% Tween®20 at 4°C with gentle shaking, overnight. subsidiaries. Tween is a registered trademark of ICI Americas, Inc. Applications Key: W—Western IP—Immunoprecipitation IHC—Immunohistochemistry ChIP—Chromatin Immunoprecipitation IF—Immunofluorescence F—Flow cytometry E-P—ELISA-Peptide Species Cross-Reactivity Key: H—human M—mouse R—rat Hm—hamster Mk—monkey Mi—mink C—chicken Dm—D. melanogaster X—Xenopus Z—zebrafish B—bovine ® 2015 Cell Signaling Technology, Inc. ® 2015 Cell Signaling Technology, Inc. are trademarks of Cell Signaling Technology, XP and Cell Signaling Technology Dg—dog Pg—pig Sc—S. cerevisiae Ce—C. elegans Hr—horse All—all species expected Species enclosed in parentheses are predicted to react based on 100% homology..