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Lieferung: frei Haus Bestellung auf Rechnung SZABO-SCANDIC Lieferung: € 10,- HandelsgmbH & Co KG Erstbestellung Vorauskassa Quellenstraße 110, A-1100 Wien T. +43(0)1 489 3961-0 Zuschläge F. +43(0)1 489 3961-7 [email protected] • Mindermengenzuschlag www.szabo-scandic.com • Trockeneiszuschlag • Gefahrgutzuschlag linkedin.com/company/szaboscandic • Expressversand facebook.com/szaboscandic SANTA CRUZ BIOTECHNOLOGY, INC. ICMT siRNA (m): sc-146137

BACKGROUND CHROMOSOMAL LOCATION ICMT (isoprenylcysteine carboxylmethyltransferase), also known as PCCMT, Genetic locus: Icmt (mouse) mapping to 4 E2. HSTE14 or PPMT, is a 284 amino acid multi-pass membrane protein that localizes to the endoplasmic reticulum and belongs to the isoprenylcysteine PRODUCT O-methyltransferase family. Expressed ubiquitously with highest expression ICMT siRNA (m) is a pool of 3 target-specific 19-25 nt siRNAs designed to in brain tissue, ICMT uses divalent cations (such as zinc) to catalyze the post- knock down gene expression. Each vial contains 3.3 nmol of lyophilized translational methylation of isoprenylated C-terminal cysteine residues. siRNA, sufficient for a 10 µM solution once resuspended using protocol Specifically, ICMT functions as the third of three enzymes that modify iso- below. Suitable for 50-100 transfections. Also see ICMT shRNA Plasmid (m): prenylated C-terminal cysteine residues on target proteins, thereby targeting sc-146137-SH and ICMT shRNA (m) Lentiviral Particles: sc-146137-V as those proteins to the cell membrane. Inhibition of ICMT effects Ras function alternate gene silencing products. and causes endothelial apoptosis, an event that may lead to a decrease in oncogenic transformation. Multiple isoforms of ICMT exist due to alternative For independent verification of ICMT (m) gene silencing results, we splicing. also provide the individual siRNA duplex components. Each is available as 3.3 nmol of lyophilized siRNA. These include: sc-146137A, sc-146137B and REFERENCES sc-146137C. 1. Dai, Q., Choy, E., Chiu, V., Romano, J., Slivka, S.R., Steitz, S.A., Michaelis, S. STORAGE AND RESUSPENSION and Philips, M.R. 1998. Mammalian prenylcysteine carboxyl methyltrans- ferase is in the endoplasmic reticulum. J. Biol. Chem. 273: 15030-15034. Store lyophilized siRNA duplex at -20° C with desiccant. Stable for at least one year from the date of shipment. Once resuspended, store at -20° C, 2. Desrosiers, R.R., Nguyen, Q.T. and Beliveau, R. 1999. The carboxyl methyl- avoid contact with RNAses and repeated freeze thaw cycles. transferase modifying G proteins is a metalloenzyme. Biochem. Biophys. Res. Commun. 261: 790-797. Resuspend lyophilized siRNA duplex in 330 µl of the RNAse-free water provided. Resuspension of the siRNA duplex in 330 µl of RNAse-free water 3. Choy, E., Chiu, V.K., Silletti, J., Feoktistov, M., Morimoto, T., Michaelson, D., makes a 10 µM solution in a 10 µM Tris-HCl, pH 8.0, 20 mM NaCl, 1 mM Ivanov, I.E. and Philips, M.R. 1999. Endomembrane trafficking of Ras: the EDTA buffered solution. CAAX motif targets proteins to the ER and Golgi. Cell 98: 69-80. 4. Online Mendelian Inheritance in Man, OMIM™. 2002. Johns Hopkins APPLICATIONS University, Baltimore, MD. MIM Number: 605851. World Wide Web URL: ICMT siRNA (m) is recommended for the inhibition of ICMT expression in http://www.ncbi.nlm.nih.gov/omim/ mouse cells. 5. Kramer, K., Harrington, E.O., Lu, Q., Bellas, R., Newton, J., Sheahan, K.L. and Rounds, S. 2003. Isoprenylcysteine carboxyl methyltransferase activity SUPPORT REAGENTS modulates endothelial cell apoptosis. Mol. Biol. Cell 14: 848-857. For optimal siRNA transfection efficiency, Santa Cruz Biotechnology’s 6. Winter-Vann, A.M., Kamen, B.A., Bergo, M.O., Young, S.G., Melnyk, S., siRNA Transfection Reagent: sc-29528 (0.3 ml), siRNA Transfection Medium: James, S.J. and Casey, P.J. 2003. Targeting Ras signaling through inhibi- sc-36868 (20 ml) and siRNA Dilution Buffer: sc-29527 (1.5 ml) are recom- tion of carboxyl methylation: an unexpected property of methotrexate. mended. Control siRNAs or Fluorescein Conjugated Control siRNAs are Proc. Natl. Acad. Sci. USA 100: 6529-6534. available as 10 µM in 66 µl. Each contain a scrambled sequence that will not lead to the specific degradation of any known cellular mRNA. Fluorescein 7. Bergo, M.O., Gavino, B.J., Hong, C., Beigneux, A.P., McMahon, M., Casey, Conjugated Control siRNAs include: sc-36869, sc-44239, sc-44240 and P.J. and Young, S.G. 2004. Inactivation of Icmt inhibits transformation by sc-44241. Control siRNAs include: sc-37007, sc-44230, sc-44231, sc-44232, oncogenic K-Ras and B-Raf. J. Clin. Invest. 113: 539-550. sc-44233, sc-44234, sc-44235, sc-44236, sc-44237 and sc-44238. 8. Roberts, M.J., Troutman, J.M., Chehade, K.A., Cha, H.C., Kao, J.P., Huang, X., Zhan, C.G., Peterson, Y.K., Subramanian, T., Kamalakkannan, S., Andres, RT-PCR REAGENTS D.A. and Spielmann, H.P. 2006. Hydrophilic anilinogeranyl diphosphate Semi-quantitative RT-PCR may be performed to monitor ICMT gene expres- prenyl analogues are Ras function inhibitors. Biochemistry 45: 15862-15872. sion knockdown using RT-PCR Primer: ICMT (m)-PR: sc-146137-PR (20 µl). 9. Lu, Q., Harrington, E.O., Newton, J., Jankowich, M. and Rounds, S. 2007. Annealing temperature for the primers should be 55-60° C and the extension Inhibition of ICMT induces endothelial cell apoptosis through GRP94. Am. temperature should be 68-72° C. J. Respir. Cell Mol. Biol. 37: 20-30. RESEARCH USE PROTOCOLS For research use only, not for use in diagnostic procedures. See our web site at www.scbt.com for detailed protocols and support products.

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