SANTA CRUZ BIOTECHNOLOGY, INC. ECOP shRNA (h) Lentiviral Particles: sc-89730-V BACKGROUND APPLICATIONS ECOP (EGFR-coamplified and overexpressed protein), also known as VOPP1 ECOP shRNA (h) Lentiviral Particles is recommended for the inhibition of (vesicular, overexpressed in cancer, prosurvival protein 1) or GASP (glioblas- ECOP expression in human cells. toma-amplified secreted protein), is a 172 amino acid protein that is coam- plified with EGFR and overexpressed in multiple glioblastomas. Highly ex- SUPPORT REAGENTS pressed in ovary and thymus, ECOP is found at moderate levels in testis, Control shRNA Lentiviral Particles: sc-108080. Available as 200 µl frozen colon, small intestine, and spleen, and at low levels in liver, placenta and viral stock containing 1.0 x 106 infectious units of virus (IFU); contains an κ prostate. ECOP regulates NF B signaling and may have a role in resistance shRNA construct encoding a scrambled sequence that will not lead to the to apoptosis. The gene encoding ECOP maps to human chromosome 7, which specific degradation of any known cellular mRNA. houses over 1,000 genes, comprises nearly 5% of the human genome and has been linked to Osteogenesis imperfecta, Pendred syndrome, Lissencephaly, RT-PCR REAGENTS Citrullinemia and Shwachman-Diamond syndrome. Semi-quantitative RT-PCR may be performed to monitor ECOP gene expres- REFERENCES sion knockdown using RT-PCR Primer: ECOP (h)-PR: sc-89730-PR (20 µl). Annealing temperature for the primers should be 55-60° C and the extension 1. Tsipouras, P., Myers, J.C., Ramirez, F. and Prockop, D.J. 1983. Restriction temperature should be 68-72° C. fragment length polymorphism associated with the pro α 2(I) gene of human type I procollagen. Application to a family with an autosomal dom- BIOSAFETY inant form of osteogenesis imperfecta. J. Clin. Invest. 72: 1262-1267. Lentiviral particles can be employed in standard Biosafety Level 2 tissue 2. Collins, V.P. 1995. Gene amplification in human gliomas. Glia 15: 289-296. culture facilities (and should be treated with the same level of caution as 3. Iwasaki, S., Usami, S., Abe, S., Isoda, H., Watanabe, T. and Hoshino, T. with any other potentially infectious reagent). Lentiviral particles are repli- 2001. Long-term audiological feature in Pendred syndrome caused by PDS cation-incompetent and are designed to self-inactivate after transduction mutation. Arch. Otolaryngol. Head Neck Surg. 127: 705-708. and integration of shRNA constructs into genomic DNA of target cells. 4. Eley, G.D., Reiter, J.L., Pandita, A., Park, S., Jenkins, R.B., Maihle, N.J. and RESEARCH USE James, C.D. 2002. A chromosomal region 7p11.2 transcript map: its devel- opment and application to the study of EGFR amplicons in glioblastoma. The purchase of this product conveys to the buyer the nontransferable right Neurooncology 4: 86-94. to use the purchased amount of the product and all replicates and derivatives for research purposes conducted by the buyer in his laboratory only (whether 5. Park, S. and James, C.D. 2005. ECop (EGFR-coamplified and overexpressed the buyer is an academic or for-profit entity). The buyer cannot sell or other- protein), a novel protein, regulates NFκB transcriptional activity and asso- wise transfer (a) this product (b) its components or (c) materials made using ciated apoptotic response in an IκBα-dependent manner. Oncogene 24: this product or its components to a third party, or otherwise use this product 2495-2502. or its components or materials made using this product or its components for Commercial Purposes. CHROMOSOMAL LOCATION Genetic locus: VOPP1 (human) mapping to 7p11.2. PROTOCOLS See our web site at www.scbt.com for detailed protocols and support PRODUCT products. ECOP shRNA (h) Lentiviral Particles are concentrated, transduction-ready viral particles containing a target-specific construct that encodes a 19-25 nt (plus hairpin) shRNA designed to knock down gene expression. Each vial contains 200 µl frozen stock containing 1.0 x 106 infectious units of virus (IFU) in Dulbecco’s Modified Eagle’s Medium with 25 mM HEPES pH 7.3. Suitable for 10-20 transductions. Also see ECOP siRNA (h): sc-89730 and ECOP shRNA Plasmid (h): sc-89730-SH as alternate gene silencing products. STORAGE Store lentiviral particles at -80º C. Stable for at least one year from the date of shipment. Once thawed, particles can be stored at 4º C for up to one week. Avoid repeated freeze thaw cycles. Santa Cruz Biotechnology, Inc. 1.800.457.3801 831.457.3800 fax 831.457.3801 Europe +00800 4573 8000 49 6221 4503 0 www.scbt.com.