SANTA CRUZ BIOTECHNOLOGY, INC. VMP1 shRNA Plasmid (m): sc-154473-SH

BACKGROUND CHROMOSOMAL LOCATION TMEM49 (transmembrane 49), also known as TDC1 or VMP1 (vacuole Genetic locus: Vmp1 (mouse) mapping to 11 C. membrane protein 1), is a 406 amino acid multi-pass membrane protein that localizes to the endoplasmic reticulum-Golgi intermediate compartment mem- PRODUCT brane. Overexpression of TMEM49, a stress-induced protein, results in the VMP1 shRNA Plasmid (m) is a pool of 3 target-specific lentiviral vector formation of intracellular vacuoles followed by cell death, suggesting that plasmids each encoding 19-25 nt (plus hairpin) shRNAs designed to knock TMEM49 plays an important role in the maintenance of cellular integrity. down expression. Each plasmid contains a puromycin resistance gene Additionally, TMEM49 may be involved in the early stages of acute pancre- for the selection of cells stably expressing shRNA. Each vial contains 20 µg atitis. The gene encoding TMEM49 maps to 17, which compris- of lyophilized shRNA plasmid DNA. Suitable for up to 20 transfections. Also es over 2.5% of the and encodes over 1,200 . Two key see VMP1 siRNA (m): sc-154473 and VMP1 shRNA (m) Lentiviral Particles: tumor suppressor genes are associated with , namely, p53 sc-154473-V as alternate gene silencing products. and BRCA1. Tumor suppressor p53 is necessary for maintenance of cellular genetic integrity by moderating cell fate through DNA repair versus cell death. STORAGE AND RESUSPENSION Malfunction or loss of p53 expression is associated with malignant cell growth and Li-Fraumeni syndrome. Like p53, BRCA1 is directly involved in Store lyophilized shRNA plasmid DNA at 4° C with desiccant. Stable for DNA repair, though specifically it is recognized as a genetic determinant of at least one year from the date of shipment. Once resuspended, store at early onset breast cancer and predisposition to cancers of the ovary, colon, 4° C for short term storage or -80° C for long term storage. Avoid repeated prostate gland and fallopian tubes. freeze thaw cycles. Resuspend lyophilized shRNA plasmid DNA in 200 µl of the deionized water REFERENCES provided. Resuspension of the shRNA plasmid DNA in 200 µl of deionized 1. Dusetti, N.J., Jiang, Y., Vaccaro, M.I., Tomasini, R., Azizi Samir, A., Calvo, water makes a 0.1 µg/µl solution in a 10 mM Tris, 1 mM EDTA buffered E.L., Ropolo, A., Fiedler, F., Mallo, G.V., Dagorn, J.C. and Iovanna, J.L. 2002. solution. Cloning and expression of the rat vacuole membrane protein 1 (VMP1), a new gene activated in pancreas with acute pancreatitis, which promotes APPLICATIONS vacuole formation. Biochem. Biophys. Res. Commun. 290: 641-649. VMP1 shRNA Plasmid (m) is recommended for the inhibition of VMP1 2. Online Mendelian Inheritance in Man, OMIM™. 2002. Johns Hopkins expression in mouse cells. University, Baltimore, MD. MIM Number: 611753. World Wide Web URL: http://www.ncbi.nlm.nih.gov/omim/ SUPPORT REAGENTS For optimal shRNA Plasmid transfection efficiency, Santa Cruz Biotechnology’s 3. Nusbaum, R., Vogel, K.J. and Ready, K. 2006-2007. Susceptibility to breast shRNA Plasmid Transfection Reagent: sc-108061 (0.2 ml) and shRNA Plasmid cancer: hereditary syndromes and low penetrance genes. Breast Dis. 27: Transfection Medium: sc-108062 (20 ml) are recommended. Control shRNAs 21-50. are available as 20 µg lyophilized plasmid DNA. Each encodes a scrambled 4. Ropolo, A., Grasso, D., Pardo, R., Sacchetti, M.L., Archange, C., Lo Re, A., shRNA sequence that will not lead to the specific degradation of any known Seux, M., Nowak, J., Gonzalez, C.D., Iovanna, J.L. and Vaccaro, M.I. 2007. cellular mRNA. Control shRNA Plasmids include: sc-108060, sc-108065 and The pancreatitis-induced vacuole membrane protein 1 triggers autophagy sc-108066. in mammalian cells. J. Biol. Chem. 282: 37124-37133. 5. Tai, Y.C., Domchek, S., Parmigiani, G. and Chen, S. 2007. Breast cancer RT-PCR REAGENTS risk among male BRCA1 and BRCA2 mutation carriers. J. Natl. Cancer Semi-quantitative RT-PCR may be performed to monitor VMP1 gene Inst. 99: 1811-1814. expression knockdown using RT-PCR Primer: VMP1 (m)-PR: sc-154473-PR (20 µl). Annealing temperature for the primers should be 55-60° C and the 6. Yan, J., Jiang, J., Lim, C.A., Wu, Q., Ng, H.H. and Chin, K.C. 2007. BLIMP1 extension temperature should be 68-72° C. regulates cell growth through repression of p53 transcription. Proc. Natl. Acad. Sci. USA 104: 1841-1846. RESEARCH USE 7. Sauermann, M., Sahin, O., Sültmann, H., Hahne, F., Blaszkiewicz, S., The purchase of this product conveys to the buyer the nontransferable right Majety, M., Zatloukal, K., Füzesi, L., Poustka, A., Wiemann, S. and Arlt, D. to use the purchased amount of the product and all replicates and derivatives 2008. Reduced expression of vacuole membrane protein 1 affects the for research purposes conducted by the buyer in his laboratory only (whether invasion capacity of tumor cells. Oncogene 27: 1320-1326. the buyer is an academic or for-profit entity). The buyer cannot sell or other- wise transfer (a) this product (b) its components or (c) materials made using PROTOCOLS this product or its components to a third party, or otherwise use this product See our web site at www.scbt.com for detailed protocols and support or its components or materials made using this product or its components products. for Commercial Purposes.

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