SAN TA C RUZ BI OTEC HNOL OG Y, INC . Muskelin shRNA (m) Lentiviral Particles: sc-149721-V

BACKGROUND APPLICATIONS Muskelin, also designated MKLN1 or TWA2, is a 735 amino acid nuclear and Muskelin shRNA (m) Lentiviral Particles is recommended for the inhibition cytoplasmic that mediates cytoskeletal responses and cell spreading of Muskelin expression in mouse cells. to Thrombospondin 1, an extracellular matrix glycoprotein. Forming a CTLH complex with RMND5A, Ran BP-M and TWA1, Muskelin exists as two alterna - SUPPORT REAGENTS tively spliced isoforms. Muskelin is implicated as an isoform-specific anchoring Control shRNA Lentiviral Particles: sc-108080. Available as 200 µl frozen protein for the prostaglandin EP3 receptor, and contains one LisH domain, viral stock containing 1.0 x 10 6 infectious units of virus (IFU); contains an a CTLH domain, six Kelch repeats and an N-terminal discoidin-like domain shRNA construct encoding a scrambled sequence that will not lead to the through which it self-associates via a head-to-tail mechanism. Human and specific degradation of any known cellular mRNA. mouse Muskelin share 98% amino acid seqeuence homology and are encoded by located on 7q32.3 and 6 A3.3, respectively. EXPRESSION MONITORING REFERENCES Muskelin (C-12): sc-398956 is recommended as a control antibody for moni - toring of Muskelin knockdown by Western Blotting (starting 1. Adams, J.C., et al. 1998. Muskelin, a novel intracellular mediator of cell dilution 1:200, dilution range 1:100-1:1000) or immunofluorescence (starting adhesive and cytoskeletal responses to thrombospondin-1. EMBO J. 17: dilution 1:50, dilution range 1:50-1:500). 4964-4974. 2. Adams, J.C., et al. 1999. cDNA cloning of human muskelin and localisa - RT-PCR REAGENTS tion of the muskelin (MKLN1) gene to human 7q32 and Semi-quantitative RT-PCR may be performed to monitor Muskelin gene mouse chromosome 6 B1/B2 by physical mapping and FISH. Cytogenet. expression knockdown using RT-PCR Primer: Muskelin (m)-PR: sc-149721-PR Cell Genet. 87: 19-21. (20 µl). Annealing temperature for the primers should be 55-60° C and the 3. Hasegawa, H., et al. 2000. Receptor isoform-specific interaction of extension temperature should be 68-72° C. prostaglandin EP3 receptor with muskelin. Biochem. Biophys. Res. Commun. 276: 350-354. BIOSAFETY 4. Umeda, M., et al. 2003. A novel nuclear protein, Twa1, and Muskelin com - Lentiviral particles can be employed in standard Biosafety Level 2 tissue prise a complex with RanBPM. Gene 303: 47-54. culture facilities (and should be treated with the same level of caution as with any other potentially infectious reagent). Lentiviral particles are repli- 5. Prag, S., et al. 2004. Molecular analysis of muskelin identifies a conserved cation-incompetent and are designed to self-inactivate after transduction discoidin-like domain that contributes to protein self-association. Biochem. and integration of shRNA constructs into genomic DNA of target cells. J. 381: 547-559. 6. Kobayashi, N., et al. 2007. RanBPM, Muskelin, p48EMLP, p44CTLH, and RESEARCH USE the armadillo-repeat ARMC8 α and ARMC8 β are components of The purchase of this product conveys to the buyer the nontransferable right the CTLH complex. Gene 396: 236-247. to use the purchased amount of the product and all replicates and derivatives for research purposes conducted by the buyer in his laboratory only (whether CHROMOSOMAL LOCATION the buyer is an academic or for-profit entity). The buyer cannot sell or other - Genetic : Mkln1 (mouse) mapping to 6 A3.3. wise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party, or otherwise use this product PRODUCT or its components or materials made using this product or its components for Commercial Purposes. Muskelin shRNA (m) Lentiviral Particles is a pool of concentrated, transduc - tion-ready viral particles containing 3 target-specific constructs that encode PROTOCOLS 19-25 nt (plus hairpin) shRNA designed to knock down gene expression. Each vial contains 200 µl frozen stock containing 1.0 x 10 6 infectious units of virus See our web site at www.scbt.com for detailed protocols and support (IFU) in Dulbecco’s Modified Eagle’s Medium with 25 mM HEPES pH 7.3. Suit- products. able for 10-20 transductions. Also see Muskelin siRNA (m): sc-149721 and Muskelin shRNA Plasmid (m): sc-149721-SH as alternate gene silencing products.

STORAGE Store lentiviral particles at -80º C. Stable for at least one year from the date of shipment. Once thawed, particles can be stored at 4º C for up to 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