SANTA CRUZ BIOTECHNOLOGY, INC. RBM12 shRNA (h) Lentiviral Particles: sc-76364-V

BACKGROUND APPLICATIONS containing RNA recognition motifs, including various hnRNP proteins, RBM12 shRNA (h) Lentiviral Particles is recommended for the inhibition of are implicated in the regulation of alternative splicing and components RBM12 expression in human cells. of snRNPs. The RBM (RNA-binding motif) family encodes proteins with an RNA binding motif that have been suggested to play a role in the modu- SUPPORT REAGENTS lation of apoptosis. RBM12 (RNA binding motif protein 12), also known as Control shRNA Lentiviral Particles: sc-108080. Available as 200 µl frozen SWAN, HRIHFB2091 or KIAA0765, is a 932 amino acid protein which local- viral stock containing 1.0 x 106 infectious units of virus (IFU); contains an izes to the nucleus. RBM12 contains multiple proline-rich regions, transmem- shRNA construct encoding a scrambled sequence that will not lead to the brane domains and three RNA recognition motifs (RRM). It has been found specific degradation of any known cellular mRNA. that the for RBM12 and copine I (CPNE1) overlap at human location 20q11.22, sharing the promoter region and a 5'UTR (which are con- GENE EXPRESSION MONITORING served in human, zebrafish and mouse), suggesting that a functional interac- tion between the two genes may exist. RBM12 (B-12): sc-514258 is recommended as a control antibody for moni- toring of RBM12 gene expression knockdown by Western Blotting (starting REFERENCES dilution 1:200, dilution range 1:100-1:1000) or immunofluorescence (starting dilution 1:50, dilution range 1:50-1:500). 1. Nagase, T., Ishikawa, K., Suyama, M., Kikuno, R., Miyajima, N., Tanaka, A., Kotani, H., Nomura, N. and Ohara, O. 1998. Prediction of the coding To ensure optimal results, the following support reagents are recommended: sequences of unidentified human genes. XI. The complete sequences of 1) Western Blotting: use m-IgGκ BP-HRP: sc-516102 or m-IgGκ BP-HRP (Cruz 100 new cDNA clones from brain which code for large proteins in vitro. Marker): sc-516102-CM (dilution range: 1:1000-1:10000), Cruz Marker™ DNA Res. 5: 277-286. Molecular Weight Standards: sc-2035, UltraCruz® Blocking Reagent: sc-516214 and Western Blotting Luminol Reagent: sc-2048. 2) Immunofluo- 2. Stover, C., Gradl, G., Jentsch, I., Speicher, M.R., Wieser, R. and Schwaeble, rescence: use m-IgGκ BP-FITC: sc-516140 or m-IgGκ BP-PE: sc-516141 W. 2001. cDNA cloning, chromosome assignment, and genomic structure (dilution range: 1:50-1:200) with UltraCruz® Mounting Medium: sc-24941 of a human gene encoding a novel member of the RBM family. Cytogenet. or UltraCruz® Hard-set Mounting Medium: sc-359850. Cell Genet. 92: 225-230. 3. Online Mendelian Inheritance in Man, OMIM™. 2002. Johns Hopkins RT-PCR REAGENTS University, Baltimore, MD. MIM Number: 607179. World Wide Web URL: Semi-quantitative RT-PCR may be performed to monitor RBM12 gene expres- http://www.ncbi.nlm.nih.gov/omim/ sion knockdown using RT-PCR Primer: RBM12 (h)-PR: sc-76364-PR (20 µl). 4. Maris, C., Dominguez, C. and Allain, F.H. 2005. The RNA recognition motif, Annealing temperature for the primers should be 55-60° C and the extension a plastic RNA-binding platform to regulate post-transcriptional gene expres- temperature should be 68-72° C. sion. FEBS J. 272: 2118-2131. BIOSAFETY 5. Yang, W., Ng, P., Zhao, M., Wong, T.K., Yiu, S.M. and Lau, Y.L. 2008. Promoter-sharing by different genes in —CPNE1 and Lentiviral particles can be employed in standard Biosafety Level 2 tissue RBM12 gene pair as an example. BMC Genomics 9: 456. culture facilities (and should be treated with the same level of caution as with any other potentially infectious reagent). Lentiviral particles are repli- CHROMOSOMAL LOCATION cation-incompetent and are designed to self-inactivate after transduction and integration of shRNA constructs into genomic DNA of target cells. Genetic locus: RBM12 (human) mapping to 20q11.22. RESEARCH USE PRODUCT The purchase of this product conveys to the buyer the nontransferable right RBM12 shRNA (h) Lentiviral Particles is a pool of concentrated, transduction- to use the purchased amount of the product and all replicates and derivatives ready viral particles containing 3 target-specific constructs that encode 19- for research purposes conducted by the buyer in his laboratory only (whether 25 nt (plus hairpin) shRNA designed to knock down gene expression. Each the buyer is an academic or for-profit entity). The buyer cannot sell or other- 6 vial contains 200 µl frozen stock containing 1.0 x 10 infectious units of virus wise transfer (a) this product (b) its components or (c) materials made using (IFU) in Dulbecco’s Modified Eagle’s Medium with 25 mM HEPES pH 7.3. this product or its components to a third party, or otherwise use this product Suitable for 10-20 transductions. Also see RBM12 siRNA (h): sc-76364 and or its components or materials made using this product or its components RBM12 shRNA Plasmid (h): sc-76364-SH as alternate gene silencing products. for Commercial Purposes.

STORAGE PROTOCOLS Store lentiviral particles at -80º C. Stable for at least one year from the See our web site at www.scbt.com for detailed protocols and support date of shipment. Once thawed, particles can be stored at 4º C for up to products. 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