SANTA CRUZ BIOTECHNOLOGY, INC. POLR2L shRNA Plasmid (h): sc-96685-SH

BACKGROUND CHROMOSOMAL LOCATION POLR2L (polymerase (RNA) II (DNA directed) polypeptide L, 7.6 kDa), also Genetic locus: POLR2L (human) mapping to 11p15.5. known as RBP10, RPB10, RPABC5, RPB7.6, hsRPB10b or RPB10β, is a 67 amino acid that belongs to the archaeal rpoN/eukaryotic RPB10 RNA PRODUCT polymerase subunit family. Localizing to nucleus, POLR2L comprises two POLR2L shRNA Plasmid (h) is a pool of 2 target-specific lentiviral vector exons, interspaced with an intron of approximately 2.1 kb. Containing four plasmids each encoding 19-25 nt (plus hairpin) shRNAs designed to knock conserved cysteines characteristic of an atypical zinc-binding domain, down expression. Each plasmid contains a puromycin resistance gene POLR2L encodes a subunit of RNA polymerase II, which synthesizes mRNA for the selection of cells stably expressing shRNA. Each vial contains 20 µg in eukaryotes. POLR2L participates in transcription regulation, DNA-directed of lyophilized shRNA plasmid DNA. Suitable for up to 20 transfections. Also RNA polymerase activity, DNA binding and zinc ion binding. POLR2L is a see POLR2L siRNA (h): sc-96685 and POLR2L shRNA (h) Lentiviral Particles: component of the RNA polymerase I (Pol I), RNA polymerase II (Pol II) and sc-96685-V as alternate gene silencing products. RNA polymerase III (Pol III) complexes, which consist of at least 13, 12 and 17 subunits, respectively. The gene that encodes POLR2L maps to human STORAGE AND RESUSPENSION 11p15.5. Store lyophilized shRNA plasmid DNA at 4° C with desiccant. Stable for REFERENCES at least one year from the date of shipment. Once resuspended, store at 4° C for short term storage or -80° C for long term storage. Avoid repeated 1. Acker, J., et al. 1996. The gene (POLR2L) encoding the hRPB7.6 subunit of freeze thaw cycles. human RNA polymerase. Genomics 32: 86-90. Resuspend lyophilized shRNA plasmid DNA in 200 µl of the deionized water 2. Rubie, C., et al. 2005. variability in normal and cancer- provided. Resuspension of the shRNA plasmid DNA in 200 µl of deionized ous colorectal, pancreatic, esophageal, gastric and hepatic tissues. Mol. water makes a 0.1 µg/µl solution in a 10 mM Tris, 1 mM EDTA buffered Cell. Probes. 19: 101-109. solution. 3. Boucneau, J., et al. 2005. profiling of cultured human NF1 heterozygous (NF1+/-) melanocytes reveals downregulation of a transcrip- APPLICATIONS tional cis-regulatory network mediating activation of the melanocyte-spe- POLR2L shRNA Plasmid (h) is recommended for the inhibition of POLR2L cific dopachrome tautomerase (DCT) gene. Pigment Cell Res. 18: 285-299. expression in human cells. 4. Bermúdez-Soto, M.J., et al. 2007. Up-regulation of tumor suppressor carci- noembryonic antigen-related cell adhesion molecule 1 in human colon SUPPORT REAGENTS cancer Caco-2 cells following repetitive exposure to dietary levels of a For optimal shRNA Plasmid transfection efficiency, Santa Cruz Biotechnology’s polyphenol-rich chokeberry juice. J. Nutr. Biochem. 18: 259-271. shRNA Plasmid Transfection Reagent: sc-108061 (0.2 ml) and shRNA Plasmid 5. Zhou, H., et al. 2008. Genome-scale RNAi screen for host factors required Transfection Medium: sc-108062 (20 ml) are recommended. Control shRNAs for HIV replication. Cell Host Microbe 4: 495-504. 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 6. Romanowski, T., et al. 2008. GUS and PMM1 as suitable reference cellular mRNA. Control shRNA Plasmids include: sc-108060, sc-108065 and for gene expression analysis in the liver tissue of patients with chronic sc-108066. hepatitis. Med. Sci. Monit. 14: BR147-BR152. 7. Ho-Pun-Cheung, A., et al. 2009. Validation of an appropriate reference RT-PCR REAGENTS gene for normalization of reverse transcription-quantitative polymerase Semi-quantitative RT-PCR may be performed to monitor POLR2L gene chain reaction data from rectal cancer biopsies. Anal. Biochem. 388: expression knockdown using RT-PCR Primer: POLR2L (h)-PR: sc-96685-PR 348-350. (20 µl). Annealing temperature for the primers should be 55-60° C and the 8. Ho-Pun-Cheung, A., et al. 2009. Reverse transcription-quantitative poly- extension temperature should be 68-72° C. merase chain reaction: description of a RIN-based algorithm for accurate data normalization. BMC Mol. Biol. 10: 31. RESEARCH USE The purchase of this product conveys to the buyer the nontransferable right 9. Jantus Lewintre, E., et al. 2009. Analysis of chronic lymphotic leukemia to use the purchased amount of the product and all replicates and derivatives transcriptomic profile: differences between molecular subgroups. Leuk. for research purposes conducted by the buyer in his laboratory only (whether Lymphoma 50: 68-79. 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.

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