SANTA CRUZ BIOTECHNOLOGY, INC. ATP5D shRNA (m) Lentiviral Particles: sc-141346-V

BACKGROUND APPLICATIONS Mitochondrial ATP synthase is composed of two multi-subunit complexes ATP5D shRNA (m) Lentiviral Particles is recommended for the inhibition of that utilize an inner membrane to catalyze the syn- ATP5D expression in mouse cells. thesis of ATP during oxidative phosphorylation. The two multi-subunit com- plexes are designated F1 and F0, the former of which comprises the soluble SUPPORT REAGENTS catalytic core and the latter of which comprises the membrane-spanning Control shRNA Lentiviral Particles: sc-108080. Available as 200 µl frozen proton channel of ATP synthase. F1 consists of five distinct subunits, desig- viral stock containing 1.0 x 106 infectious units of virus (IFU); contains an nated ATP5A, ATP5B, ATP5C1, ATP5D and ATP5E, while F0 consists of ten shRNA construct encoding a scrambled sequence that will not lead to the subunits, designated ATP5H, ATP5G1, ATP5I, ATP5G2, ATP5J2, ATP5J, specific degradation of any known cellular mRNA. ATP5G3, ATP5S, ATP5F1 and ATP5L. ATP5D (ATP synthase, H+ transporting, δ mitochondrial F1 complex, subunit) is a 168 that local- RT-PCR REAGENTS izes to the mitochondrial inner membrane and is encoded by a that maps to human 19p13.3. Semi-quantitative RT-PCR may be performed to monitor ATP5D gene expres- sion knockdown using RT-PCR Primer: ATP5D (m)-PR: sc-141346-PR (20 µl). REFERENCES Annealing temperature for the primers should be 55-60° C and the extension temperature should be 68-72° C. 1. Jordan, E.M., et al. 1992. Molecular cloning of an import precursor of the δ -subunit of the human mitochondrial ATP synthase complex. Biochim. BIOSAFETY Biophys. Acta 1130: 123-126. Lentiviral particles can be employed in standard Biosafety Level 2 tissue 2. Aggeler, R., et al. 2002. A functionally active human F1F0 ATPase can be culture facilities (and should be treated with the same level of caution as purified by immunocapture from heart tissue and fibroblast cell lines. with any other potentially infectious reagent). Lentiviral particles are repli- Subunit structure and activity studies. J. Biol. Chem. 277: 33906-33912. cation-incompetent and are designed to self-inactivate after transduction 3. Hong, S., et al. 2003. ATP synthases: insights into their motor functions and integration of shRNA constructs into genomic DNA of target cells. from sequence and structural analyses. J. Bioenerg. Biomembr. 35: 95-120. RESEARCH USE 4. Cross, R.L. 2004. Molecular motors: turning the ATP motor. Nature 427: 407-408. The purchase of this product conveys to the buyer the nontransferable right to use the purchased amount of the product and all replicates and derivatives 5. Itoh, H., et al. 2004. Mechanically driven ATP synthesis by F1-ATPase. for research purposes conducted by the buyer in his laboratory only (whether Nature 427: 465-468. the buyer is an academic or for-profit entity). The buyer cannot sell or other- 6. Online Mendelian Inheritance in Man, OMIM™. 2004. Johns Hopkins wise transfer (a) this product (b) its components or (c) materials made using University, Baltimore, MD. MIM Number: 603150. World Wide Web URL: this product or its components to a third party, or otherwise use this product http://www.ncbi.nlm.nih.gov/omim/ or its components or materials made using this product or its components for Commercial Purposes. CHROMOSOMAL LOCATION PROTOCOLS Genetic : Atp5d (mouse) mapping to 10 C1. See our web site at www.scbt.com for detailed protocols and support PRODUCT products. ATP5D shRNA (m) Lentiviral Particles is a pool of concentrated, transduc- tion-ready viral particles containing 2 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 ATP5D siRNA (m): sc-141346 and ATP5D shRNA Plasmid (m): sc-141346-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