SANTA CRUZ BIOTECHNOLOGY, INC. DDX27 shRNA Plasmid (m): sc-77412-SH

BACKGROUND STORAGE AND RESUSPENSION DDX27 (probable ATP-dependent RNA DDX27) is a 796 amino acid Store lyophilized shRNA plasmid DNA at 4° C with desiccant. Stable for encoded by the human DDX27. This protein belongs to the DEAD at least one year from the date of shipment. Once resuspended, store at box helicase family, DDX27/DRS1 subfamily and contains one helicase ATP- 4° C for short term storage or -80° C for long term storage. Avoid repeated binding domain and one helicase C-terminal domain. DDX27 is a nuclear freeze thaw cycles. protein found on 5 that likely functions as an ATP-dependent Resuspend lyophilized shRNA plasmid DNA in 200 µl of the deionized water RNA helicase. RNA are highly conserved enzymes that utilize the provided. Resuspension of the shRNA plasmid DNA in 200 µl of deionized energy derived from NTP hydrolysis to modulate the structure of RNA. RNA water makes a 0.1 µg/µl solution in a 10 mM Tris, 1 mM EDTA buffered helicases participate in all biological processes that involve RNA, including solution. transcription, splicing and translation. APPLICATIONS REFERENCES DDX27 shRNA Plasmid (m) is recommended for the inhibition of DDX27 1. O’Day, C.L., Chavanikamannil, F. and Abelson, J. 1996. 18S rRNA pro- expression in mouse cells. cessing requires the RNA helicase-like protein Rrp3. Nucleic Acids Res. 24: 3201-3207. SUPPORT REAGENTS 2. Doorbar, J., Elston, R.C., Napthine, S., Raj, K., Medcalf, E., Jackson, D., For optimal shRNA Plasmid transfection efficiency, Santa Cruz Biotechnology’s Coleman, N., Griffin, H.M., Masterson, P., Stacey, S., Mengistu, Y. and shRNA Plasmid Transfection Reagent: sc-108061 (0.2 ml) and shRNA Plasmid Dunlop, J. 2000. The E1E4 protein of human papillomavirus type 16 asso- Transfection Medium: sc-108062 (20 ml) are recommended. Control shRNAs ciates with a putative RNA helicase through sequences in its C terminus. are available as 20 µg lyophilized plasmid DNA. Each encodes a scrambled J. Virol. 74: 10081-10095. shRNA sequence that will not lead to the specific degradation of any known 3. Regard, J.B., Scheek, S., Borbiev, T., Lanahan, A.A., Schneider, A., cellular mRNA. Control shRNA Plasmids include: sc-108060, sc-108065 and Demetriades, A.M., Hiemisch, H., Barnes, C.A., Verin, A.D. and Worley, sc-108066. P.F. 2004. Verge: a novel vascular early response gene. J. Neurosci. 24: 4092-4103. MONITORING 4. Abdelhaleem, M. 2005. RNA helicases: regulators of differentiation. Clin. DDX27 (2251C2a): sc-81074 is recommended as a control for moni- Biochem. 38: 499-503. toring of DDX27 gene expression knockdown by Western Blotting (starting dilution 1:200, dilution range 1:100-1:1000) or immunofluorescence (starting 5. Lee, J.H., Rho, S.B. and Chun, T. 2005. GABAA receptor-associated pro- dilution 1:50, dilution range 1:50-1:500). tein (GABARAP) induces apoptosis by interacting with DEAD (Asp-Glu-Ala- Asp/His) box polypeptide 47 (DDX47). Biotechnol. Lett. 27: 623-628. RT-PCR REAGENTS 6. Nousiainen, M., Sillje, H.H., Sauer, G., Nigg, E.A. and Körner, R. 2006. Semi-quantitative RT-PCR may be performed to monitor DDX27 gene expres- Phosphoproteome analysis of the human mitotic spindle. Proc. Natl. sion knockdown using RT-PCR Primer: DDX27 (m)-PR: sc-77412-PR (20 µl). Acad. Sci. USA 103: 5391-5396. Annealing temperature for the primers should be 55-60° C and the extension 7. Sekiguchi, T., Hayano, T., Yanagida, M., Takahashi, N. and Nishimoto, T. temperature should be 68-72° C. 2006. NOP132 is required for proper nucleolus localization of DEAD-box RNA helicase DDX47. Nucleic Acids Res. 34: 4593-4608. RESEARCH USE The purchase of this product conveys to the buyer the nontransferable right CHROMOSOMAL LOCATION to use the purchased amount of the product and all replicates and derivatives Genetic : Ddx27 (mouse) mapping to 2 H3. for research purposes conducted by the buyer in his laboratory only (whether the buyer is an academic or for-profit entity). The buyer cannot sell or other- PRODUCT wise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party, or otherwise use this product DDX27 shRNA Plasmid (m) is a pool of 3 target-specific lentiviral vector or its components or materials made using this product or its components plasmids each encoding 19-25 nt (plus hairpin) shRNAs designed to knock for Commercial Purposes. down gene expression. Each plasmid contains a puromycin resistance gene for the selection of cells stably expressing shRNA. Each vial contains 20 µg PROTOCOLS of lyophilized shRNA plasmid DNA. Suitable for up to 20 transfections. Also see DDX27 siRNA (m): sc-77412 and DDX27 shRNA (m) Lentiviral Particles: See our web site at www.scbt.com for detailed protocols and support sc-77412-V as alternate gene silencing products. products.

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