SANTA CRUZ BIOTECHNOLOGY, INC. BinCARD shRNA Plasmid (m): sc-141706-SH

BACKGROUND PRODUCT BinCARD (Bcl10-interacting CARD ) is a 228 amino acid protein that BinCARD shRNA Plasmid (m) is a target-specific lentiviral vector plasmid exists as two alternatively spiced isoforms. BinCARD localizes to nucleus encoding a 19-25 nt (plus hairpin) shRNA designed to knock down and is expressed in ovary, testis, placenta, skeletal muscle, kidney, lung, heart, expression. Each plasmid contains a puromycin resistance gene for the liver, thymus and brain. Containing a CARD domain, BinCARD plays a role in selection of cells stably expressing shRNA. Each vial contains 20 µg of inhibiting the effects of Bcl10-induced activation of NFκB possibly by inhibit- lyophilized shRNA plasmid DNA. Suitable for up to 20 transfections. Also ing the phosphorylation of Bcl10 in a CARD-dependent manner. The BinCARD see BinCARD siRNA (m): sc-141706 and BinCARD shRNA (m) Lentiviral gene maps to 9q22.31. consists of about 145 Particles: sc-141706-V as alternate gene silencing products. million bases and 4% of the and encodes nearly 900 . Notably, chromosome 9 encompasses the largest interferon family gene clus- STORAGE AND RESUSPENSION ter. Considered to play a role in gender determination, deletion of the distal Store lyophilized shRNA plasmid DNA at 4° C with desiccant. Stable for portion of 9p can lead to development of male to female sex reversal, the at least one year from the date of shipment. Once resuspended, store at phenotype of a female with a male X,Y genotype. Hereditary hemorrhagic 4° C for short term storage or -80° C for long term storage. Avoid repeated telangiectasia, which is characterized by harmful vascular defects, is associ- freeze thaw cycles. ated with the chromosome 9 gene encoding endoglin protein, ENG. Familial dysautonomia is also associated with chromosome 9 though through the gene Resuspend lyophilized shRNA plasmid DNA in 200 µl of the deionized water IKAP. provided. Resuspension of the shRNA plasmid DNA in 200 µl of deionized water makes a 0.1 µg/µl solution in a 10 mM Tris, 1 mM EDTA buffered REFERENCES solution. 1. Strausberg, R.L., et al. 2002. Generation and initial analysis of more than APPLICATIONS 15,000 full-length human and mouse cDNA sequences. Proc. Natl. Acad. Sci. USA 99: 16899-16903. BinCARD shRNA Plasmid (m) is recommended for the inhibition of BinCARD expression in mouse cells. 2. Woo, H.N., et al. 2004. Inhibition of Bcl10-mediated activation of NFκB by BinCARD, a Bcl10-interacting CARD protein. FEBS Lett. 578: 239-244. SUPPORT REAGENTS 3. Ota, T., et al. 2004. Complete sequencing and characterization of 21,243 For optimal shRNA Plasmid transfection efficiency, Santa Cruz Biotechnology’s full-length human cDNAs. Nat. Genet. 36: 40-45. shRNA Plasmid Transfection Reagent: sc-108061 (0.2 ml) and shRNA Plasmid 4. Humphray, S.J., et al. 2004. DNA sequence and analysis of human chromo- Transfection Medium: sc-108062 (20 ml) are recommended. Control shRNAs some 9. Nature 429: 369-374. are available as 20 µg lyophilized plasmid DNA. Each encodes a scrambled shRNA sequence that will not lead to the specific degradation of any known 5. Wang, P., et al. 2006. Cloning of a novel human caspase-9 splice variant cellular mRNA. Control shRNA Plasmids include: sc-108060, sc-108065 and containing only the CARD domain. Life Sci. 79: 934-940. sc-108066. 6. Temtamy, S.A., et al. 2007. Phenotypic and cytogenetic spectrum of 9p tri- somy. Genet. Couns. 18: 29-48. RT-PCR REAGENTS 7. Fernandez-L, A., et al. 2007. Gene expression fingerprinting for human Semi-quantitative RT-PCR may be performed to monitor BinCARD gene hereditary hemorrhagic telangiectasia. Hum. Mol. Genet. 16: 1515-1533. expression knockdown using RT-PCR Primer: BinCARD (m)-PR: sc-141706-PR (20 µl). Annealing temperature for the primers should be 55-60° C and the 8. Cottin, V., et al. 2007. Pulmonary vascular manifestations of hereditary extension temperature should be 68-72° C. hemorrhagic telangiectasia (rendu-osler disease). Respiration 74: 361-378. PROTOCOLS CHROMOSOMAL LOCATION See our web site at www.scbt.com for detailed protocols and support Genetic locus: 1110007C09Rik (mouse) mapping to 13 A5. products.

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