SANTA CRUZ BIOTECHNOLOGY, INC. Gα t1 (K-20): sc-389 The Power to Question BACKGROUND APPLICATIONS Heterotrimeric G proteins function to relay information from cell surface Gα t1 (K-20) is recommended for detection of Gα t1 of mouse, rat, human receptors to intracellular effectors. Each of a very broad range of receptors and bovine origin by Western Blotting (starting dilution 1:200, dilution specifically detects an extracellular stimulus (a photon, pheromone, odorant, range 1:100-1:1000), immunoprecipitation [1–2 µg per 100–500 µg of total hormone or neurotransmitter) while the effectors (i.e. adenyl cyclase), which protein (1 ml of cell lysate)] and immunofluorescence (starting dilution act to generate one or more intracellular messengers, are less numerous. In 1:50, dilution range 1:50-1:500). mammals, G protein , and polypeptides are encoded by at least 16, 4 α β γ Suitable for use as control antibody for G siRNA (h): sc-43783 and and 7 genes, respectively. Most interest in G proteins has been focused on α t1 G siRNA (m): sc-45759. their α subunits, since these proteins bind and hydrolyze GTP and most obvi- α t1 ously regulate the activity of the best studied effectors. Four distinct classes Positive Controls: rat brain extract: sc-2392 or bovine brain extract. of Gα subunits have been identified; these include Gs, Gi, Gq and Gα 12/13. The Gi class comprises all the known α subunits that are susceptible to per- RECOMMENDED SECONDARY REAGENTS tussis toxin modifications, including Gα i-1, Gα i-2, Gα i-3, Gα o, Gα t1, Gα t2, Gα z To ensure optimal results, the following support (secondary) reagents are and Gα gust. In the well characterized visual system, photorhodopsin catalyzes recommended: 1) Western Blotting: use goat anti-rabbit IgG-HRP: sc-2004 the exchange of guanine nucleotides bound to the visual transducin Gα sub- (dilution range: 1:2000-1:100,000) or Cruz Marker™ compatible goat anti- units (Gα t1 in rod cells and Gα t2 in cone cells). rabbit IgG-HRP: sc-2030 (dilution range: 1:2000-1:5000), Cruz Marker™ Molecular Weight Standards: sc-2035, TBS Blotto A Blocking Reagent: REFERENCES sc-2333 and Western Blotting Luminol Reagent: sc-2048. 2) Immunoprecip- 1. Strathmann, M. and Simon, M.I. 1990. G Protein diversity: a distinct class itation: use Protein A/G PLUS-Agarose: sc-2003 (0.5 ml agarose/2.0 ml). of α subunits is present in vertebrates and invertebrates. Proc. Natl. Acad. 3) Immunofluorescence: use goat anti-rabbit IgG-FITC: sc-2012 (dilution Sci. USA 87: 9113-9117. range: 1:100-1:400) or goat anti-rabbit IgG-TR: sc-2780 (dilution range: 1:100-1:400) with UltraCruz™ Mounting Medium: sc-24941. 2. Simon, M.I., et al. 1991. Diversity of G proteins in signal transduction. Science 252: 802-808. DATA 3. Cali, J.J., et al. 1992. Selective tissue distribution of G protein γ subunits, including a new form of the γ subunits identified by cDNA cloning. J. Biol. AB Chem. 267: 24023-24027. 83 K - 4. McLaughlin, S.K., et al. 1992. Gustducin is a taste-cell-specific G protein closely related to the transducins. Nature 357: 563-569. 51 K - < Gα t1 5. Conklin, B.R. and Bourne, H.R. 1993. Structural elements of Gα subunits 36 K - 29 K - that interact with Gβγ, receptors, and effectors. Cell 73: 631-641. CHROMOSOMAL LOCATION Gα t1 (K-20): sc-389. Western blot analysis of Gα t1 expression in rat (A) and bovine (B) brain extracts. Genetic locus: GNAT1 (human) mapping to 3p21; Gnat1 (mouse) mapping to 9 F1. SELECT PRODUCT CITATIONS SOURCE 1. Boughter, J.D., Jr., et al. 1997. Differential expression of α-Gustducin in taste bud populations of the rat and hamster. J. Neurosci. 17: 2852-2858. Gα t1 (K-20) is an affinity purified rabbit polyclonal antibody raised against a peptide mapping within a highly divergent domain of Gαt1 of human origin. 2. Weiler, R., et al. 1998. Retinoic acid has light-adaptive effects on horizon- tal cells in the retina. Proc. Natl. Acad. Sci. USA 95: 7139-7144. PRODUCT Each vial contains 200 µg IgG in 1.0 ml of PBS with < 0.1% sodium azide and RESEARCH USE 0.1% gelatin. For research use only, not for use in diagnostic procedures. Blocking peptide available for competition studies, sc-389 P, (100 µg peptide PROTOCOLS in 0.5 ml PBS containing < 0.1% sodium azide and 0.2% BSA). See our web site at www.scbt.com or our catalog for detailed protocols STORAGE and support products. Store at 4° C, **DO NOT FREEZE**. Stable for one year from the date of shipment. Non-hazardous. No MSDS required. 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.