SANTA CRUZ BIOTECHNOLOGY, INC. CEP350 shRNA (h) Lentiviral Particles: sc-78702-V

BACKGROUND APPLICATIONS CEP350 (-associated 350) is a 3,117 amino acid protein CEP350 shRNA (h) Lentiviral Particles is recommended for the inhibition of that is required for anchoring to the and for micro- CEP350 expression in human cells. tubule network integrity. As part of a ternary complex with SAS-6 and CENP- J, CEP350 plays an essential role in growth by stabilizing a procen- SUPPORT REAGENTS α triolar seed. CEP350 modulates PPAR activity by recruiting it to discrete Control shRNA Lentiviral Particles: sc-108080. Available as 200 µl frozen subcellular compartments, therefore influencing events in fatty acid metabo- viral stock containing 1.0 x 106 infectious units of virus (IFU); contains an lism. Expressed in testis, kidney, placenta, brain, heart, skeletal muscle, pan- shRNA construct encoding a scrambled sequence that will not lead to the creas. liver and lung, CEP350 is phosphorylated during mitosis. In rare and specific degradation of any known cellular mRNA. aggressive myeloproliferative disorders, a translocation results in a Flg- FOP fusion protein, resulting in constitutive kinase activity at the centrosome. RT-PCR REAGENTS CEP350 is responsible for Flg-FOP localization to the centrosome, which leads to activation of PI 3-kinase and PLC γ1 and, consequently, inappropriate sig- Semi-quantitative RT-PCR may be performed to monitor CEP350 gene expres- naling resulting in continuous cell growth. sion knockdown using RT-PCR Primer: CEP350 (h)-PR: sc-78702-PR (20 µl). Annealing temperature for the primers should be 55-60° C and the extension REFERENCES temperature should be 68-72° C. 1. Makalowska, I., et al. 2002. Identification of six novel by experimen- BIOSAFETY tal validation of GeneMachine predicted genes. Gene 284: 203-213. Lentiviral particles can be employed in standard Biosafety Level 2 tissue 2. Andersen, J.S., et al. 2003. Proteomic characterization of the human cen- culture facilities (and should be treated with the same level of caution as trosome by protein correlation profiling. Nature 426: 570-574. with any other potentially infectious reagent). Lentiviral particles are repli- 3. Patel, H., et al. 2005. Activity and subcellular compartmentalization of per- cation-incompetent and are designed to self-inactivate after transduction oxisome proliferator-activated receptor α are altered by the centrosome- and integration of shRNA constructs into genomic DNA of target cells. associated protein CAP350. J. Cell. Sci. 118: 175-186. STORAGE 4. Yan, X., et al. 2006. A complex of two centrosomal , CAP350 and FOP, cooperates with EB1 in anchoring. Mol. Biol. Cell. 17: Store lentiviral particles at -80º C. Stable for at least one year from the 634-644. date of shipment. Once thawed, particles can be stored at 4º C for up to one week. Avoid repeated freeze thaw cycles. CHROMOSOMAL LOCATION PROTOCOLS Genetic locus: CEP350 (human) mapping to 1q25.2. See our web site at www.scbt.com for detailed protocols and support PRODUCT products. CEP350 shRNA (h) Lentiviral Particles is a pool of concentrated, transduction- ready viral particles containing 3 target-specific constructs that encode 19- 25 nt (plus hairpin) shRNA designed to knock down . 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 CEP350 siRNA (h): sc-78702 and CEP350 shRNA Plasmid (h): sc-78702-SH as alternate gene silencing products.

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