Skf 97541

Manufactured by Bio-Techne

The SKF-97541 is an enzyme-linked immunosorbent assay (ELISA) kit designed to quantitatively measure the concentration of a specific analyte in a sample. The kit includes the necessary reagents and materials to perform the assay. The core function of this product is to provide a standardized and reliable method for the detection and measurement of the target analyte.

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3 protocols using skf 97541

Behavioral Assays for Fly Sleep Modulation

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All behavioral tests were performed using individual male flies, unless otherwise indicated. Each fly was housed in a 65 × 5 mm glass tube containing 5% sucrose and 2% agar (behavior food). For amino acid supplements, the indicated amount of each amino acid was dissolved in the behavior food. For oral administration of GABA-T or GABA transporter inhibitors, 10 mM of EOS (Tokyo Chemical Industry) or 10 mg/ml of NipA (Sigma) was directly dissolved in the behavior food containing the indicated amount of threonine. For oral administration of GABA receptor agonists, 10 mg/ml of THIP (Tocris) or SKF-97541 (Tocris) stock solution was diluted into the behavior food at the indicated final concentration. Flies were fed on amino acid- and/or drug-containing behavior food in LD cycles at 25°C for 4.5 days. Locomotor activity was recorded using the DAM system (Trikinetics) and quantified by the number of infrared beam crosses per minute. Sleep bouts were defined as no activity for >5 min. Sleep parameters were analyzed using an Excel macro (Pfeiffenberger et al., 2010 (link)).

Ki Y, & Lim C. (2019). Sleep-promoting effects of threonine link amino acid metabolism in Drosophila neuron to GABAergic control of sleep drive. eLife, 8, e40593.

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Characterization of Pharmacological Compounds

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2-Bromo-α-ergocryptine methanesulfonate salt (BRM) was purchased from Sigma-Aldrich. Aprepitant was obtained from Selleck. All the other compounds tested were purchased from Tocris Biosciences, including SCH 23390 hydrochloride, rotigotine hydrochloride, sumanirole maleate, B-HT 920, Ro 10-5824 dihydrochloride, YM 202074, cinnabarinic acid, MTP, ICI 118,551 hydrochloride, GS 6201, PSB 1115, SC 19220, CP 154526, L-733060, CP 96345, TAM, DOX, A 412997 dihydrochloride, AMN 082 dihydrochloride, SKF 97541, Rac BHFF, SEW 2871, purmorphamine, (R)-(−)-α-methylhistamine dihydrobromide, methimepip dihydrobromide, VU 0155041 sodium salt, antalarmin hydrochloride, NBI 35965 hydrochloride, BQ 788 sodium salt, BAY36-7620, 3-MATIDA, MPEP hydrochloride, MRS 1754, SC 51322, SC 19220, and JTE 013.

Chen Y., Palczewska G., Masuho I., Gao S., Jin H., Dong Z., Gieser L., Brooks M.J., Kiser P.D., Kern T.S., Martemyanov K.A., Swaroop A, & Palczewski K. (2016). Synergistically acting agonists and antagonists of G protein–coupled receptors prevent photoreceptor cell degeneration. Science signaling, 9(438), ra74.

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GABA receptor binding assay

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Materials and Reagents GABA, CGP54626, and SKF97541 were purchased from Tocris Bioscience. Baclofen, 3-APPA, CGP35348, CGP7930, GS39783, and rac-BHFF were purchased from Abcam. Lipofectamine 2000 and Fluo-4 AM were from Thermo Fischer Scientific. SNAP-Lumi4-Tb, SNAP-Green, SNAP-Red, CoA-Lumi4-Tb, and Tag-Lite buffer can be purchased from Cisbio Bioassays. CoA-Alexa 488, CoA-647, and Sfp synthase were from New England Biolabs. Benzyl guanine (BG) derivative fluorophores BG-Green-LC and BG-Atto465, and CGP54626-Red were a gift from Cisbio Bioassays.

Lecat-Guillet N., Monnier C., Rovira X., Kniazeff J., Lamarque L., Zwier J.M., Trinquet E., Pin J.P, & Rondard P. (2017). FRET-Based Sensors Unravel Activation and Allosteric Modulation of the GABA_B Receptor. Cell chemical biology, 24(3).

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