Sl327, by Bio-Techne - Product details

1

PTZ-Induced Seizure Assay in Mice

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PTZ dose used in this study (40 mg/kg in young mice and 50 mg/kg in adult mice, in 0.9% saline, i.p. injection volume was 1 ml/100 g body weight) was determined as a dose at which PTZ induced seizures in 75% of naive mice. In brief, 2- and 11-wk-old mice treated with drugs (saline, LPS [1.0 mg/kg/d; Sigma-Aldrich], SL327 [selective inhibitor MEK1 and MEK2; 32 mg/kg/d; Tocris Bioscience], or LPS plus SL327, i.p. injection for 3 d [P10–12 or P70–72; see details in the figure legends]) were individually placed in a Plexiglas box, and convulsive behaviors were analyzed for 30 min after PTZ injection. The time to onset and the duration of tonic-clonic seizures were measured as previously described (Shen et al., 2012 (link)).

Shen Y., Qin H., Chen J., Mou L., He Y., Yan Y., Zhou H., Lv Y., Chen Z., Wang J, & Zhou Y.D. (2016). Postnatal activation of TLR4 in astrocytes promotes excitatory synaptogenesis in hippocampal neurons. The Journal of Cell Biology, 215(5), 719-734.

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Preparation of Amyloid-β Peptide and Kinase Inhibitors

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PDTC (Sigma-Aldrich) was dissolved in sterile water and used immediately after preparation. The kinase inhibitors 10-DEBC, FPA-124, LY294002, PD198306, PD98059, PI828, SB415286, and SL327 (Tocris) were dissolved in DMSO to 10 mM. Aβ_(1-42) was purchased from American Peptide (Sunnyvale, CA) and dissolved to 1 mg/mL in sterile water. Protein concentration of the peptide was verified by DC protein assay kit (Bio Rad). The soluble peptide was used for experiments immediately after preparation as described [13 (link)].

Liddell J.R., Lehtonen S., Duncan C., Keksa-Goldsteine V., Levonen A.L., Goldsteins G., Malm T., White A.R., Koistinaho J, & Kanninen K.M. (2016). Pyrrolidine dithiocarbamate activates the Nrf2 pathway in astrocytes. Journal of Neuroinflammation, 13, 49.

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3

Ethanol and Inhibitor Preparation

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Ethanol for injection (20% v/v) was prepared by diluting 95% ethanol in 0.9% saline. All ethanol drinking solutions (v/v) were prepared by diluting 95% ethanol (Pharmco Products Inc., Brookfield, CT, USA) with tap water. The MEK1/2 inhibitor SL327 and the p38 inhibitor SB239063 (Tocris Bioscience; Ellisville, MO, USA) were freshly suspended in 45% β-cyclodextrin on the morning of each drug testing day.

Agoglia A.E., Sharko A.C., Psilos K.E., Holstein S.E., Reid G.T, & Hodge C.W. (2015). Alcohol Alters the Activation of ERK1/2, a Functional Regulator of Binge Alcohol Drinking in Adult C57BL/6J Mice. Alcoholism, clinical and experimental research, 39(3), 463-475.

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4

Neuromodulatory Drugs in Behavioral Assays

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Rapamycin (LC Laboratories) was dissolved in a vehicle solution of 1% dimethylsulfoxide (DMSO), 5% Tween-80, and saline and injected at a dose of 40 mg/kg. PF-4708671 (Tocris) was dissolved in a vehicle solution of 17% DMSO, 10% Tween-80, and saline and injected at a dose of 50 mg/kg. For intra-amygdala infusions of PF-4708671, the drug was dissolved in 100% DMSO. SL327 (Tocris) was dissolved in a vehicle solution of 5% DMSO, 5% Tween-80 and saline and injected at a dose of 30 mg/kg. Rapamycin and PF-4708671 were injected intraperitoneally (i.p.) to a volume of 5 ml/kg relative to body weight one hour before the behavioral manipulation began; SL327 was injected 45 minutes before behavior. For intra-amygdala infusions, PF-4708671 (0.2 μl) was infused bilaterally into the basolateral nucleus of the amygdala at a rate of 0.08 μl/min, one hour before behavioral testing began. The injectors remained in the guide cannula for two minutes after the infusion. Control mice received equivalent volumes of vehicle solution. Please see Supplemental Table 1 for a complete list of drugs, vehicle preparations, and injection time points relative to behavior.

Huynh T.N., Santini E., Mojica E., Fink A.E., Hall B.S., Fetcho R.N., Grosenick L., Deisseroth K., LeDoux J.E., Liston C, & Klann E. (2017). Activation of a novel p70 S6 kinase 1-dependent intracellular cascade in the basolateral nucleus of the amygdala is required for the acquisition of extinction memory. Molecular psychiatry, 23(6), 1394-1401.

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Neuromodulatory Drugs in Behavioral Assays

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Rapamycin (LC Laboratories) was dissolved in a vehicle solution of 1% dimethylsulfoxide (DMSO), 5% Tween-80, and saline and injected at a dose of 40 mg/kg. PF-4708671 (Tocris) was dissolved in a vehicle solution of 17% DMSO, 10% Tween-80, and saline and injected at a dose of 50 mg/kg. For intra-amygdala infusions of PF-4708671, the drug was dissolved in 100% DMSO. SL327 (Tocris) was dissolved in a vehicle solution of 5% DMSO, 5% Tween-80 and saline and injected at a dose of 30 mg/kg. Rapamycin and PF-4708671 were injected intraperitoneally (i.p.) to a volume of 5 ml/kg relative to body weight one hour before the behavioral manipulation began; SL327 was injected 45 minutes before behavior. For intra-amygdala infusions, PF-4708671 (0.2 μl) was infused bilaterally into the basolateral nucleus of the amygdala at a rate of 0.08 μl/min, one hour before behavioral testing began. The injectors remained in the guide cannula for two minutes after the infusion. Control mice received equivalent volumes of vehicle solution. Please see Supplemental Table 1 for a complete list of drugs, vehicle preparations, and injection time points relative to behavior.

Huynh T.N., Santini E., Mojica E., Fink A.E., Hall B.S., Fetcho R.N., Grosenick L., Deisseroth K., LeDoux J.E., Liston C, & Klann E. (2017). Activation of a novel p70 S6 kinase 1-dependent intracellular cascade in the basolateral nucleus of the amygdala is required for the acquisition of extinction memory. Molecular psychiatry, 23(6), 1394-1401.

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6

Pharmacological Evaluation of Spatial Learning

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SL327 (Tocris) was dissolved in DMSO (16 mg/ml) and was injected intraperitoneally once daily, 30 min before the water maze experiment at a dose of 32mg/kg. The volume of a single injection was under 80 μl. Lovastatin (Mevinolin, Sigma) was prepared as previously described^{22 (link)}. Briefly, lovastatin was dissolved in ethanol (final concentration of 8%) and 1N NaOH was added to convert mevinolin to the sodium salt. The pH of the final solution (4 mg/ml) was adjusted to 7.5 with HCl. The Vehicle solution was prepared with the same procedure. Lovastatin was administered daily (subcutaneous injection, 10 mg/kg) for 3 days before the first training day of the water maze and 6 h before training every day thereafter.

Lee Y.S., Ehninger D., Zhou M., Oh J.Y., Kang M., Kwak C., Ryu H.H., Butz D., Araki T., Cai Y., Balaji J., Sano Y., Nam C.I., Kim H.K., Kaang B.K., Burger C., Neel B.G, & Silva A.J. (2014). Mechanism and treatment for the learning and memory deficits associated with mouse models of Noonan syndrome. Nature neuroscience, 17(12), 1736-1743.

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7

Evaluating MEK1/2 Inhibitor Neuroprotection

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SL327 (Tocris, Bristol, UK), a MEK1/2 inhibitor (Atkins et al. 1998) proved to be toxic when dissolved in DMSO (data not shown), and therefore, was dissolved in 100% EtOH instead. P7 C57/Bl6 mice were treated with a single i.p. injection of SL327 (Dommergues et al. 2003) at 0, 15, 30, 65 or 133 μg/g BW (n = 5 per group) 20 min prior to the start of 30 min hypoxia. Fifteen minutes after HI, brains were assessed for intensity of pERK staining.

Thei L., Rocha‐Ferreira E., Peebles D., Raivich G, & Hristova M. (2018). Extracellular signal‐regulated kinase 2 has duality in function between neuronal and astrocyte expression following neonatal hypoxic–ischaemic cerebral injury. The Journal of Physiology, 596(23), 6043-6062.

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8

Modulation of Neuronal Activity

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Clozapine N-oxide (CNO; Tocris, Bristol, United Kingdom) and Deschloroclozapine dihydrochloride (DCZ; Hellobio; Bristol, United Kingdom) were freshly prepared in saline and injected intraperitoneally (i.p.) 30 min prior to behavioral tests (CNO: 1 or 10 mg/kg, DCZ: 0.2 mg/kg). SL327 (Tocris, Bristol, United Kingdom), was administered i.p. (30 mg/kg) 15 min before CNO injection. Clomipramine (CMI; Sigma, St. Louis, USA) was added to drinking water at a concentration of 40 mg/L to reach an average dose of 20 mg/kg per day.

Lee I.B., Lee E., Han N.E., Slavuj M., Hwang J.W., Lee A., Sun T., Jeong Y., Baik J.H., Park J.Y., Choi S.Y., Kwag J, & Yoon B.J. (2024). Persistent enhancement of basolateral amygdala-dorsomedial striatum synapses causes compulsive-like behaviors in mice. Nature Communications, 15, 219.

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9

Synthesis of Enantiomeric Adrenergic Compounds

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(R,R′)-MNF and (R,R ′)-fenoterol [(R,R′)-Fen] were synthesized as described previously [11 (link)]. LY294002, API-2, SL327, U0126, AM251, H-89, protein kinase inhibitor-(14–22)-amide (PKI), O-1602, and Tocrifluor 1117 (T1117) were from Tocris Bioscience. LPI, isoproterenol (ISO) and ICI-118,551 were purchased from Sigma-Aldrich. All compounds were dissolved in DMSO and were applied to cells at a final DMSO concentration of 0.1%.

Wnorowski A., Such J., Paul R.K., Wersto R.P., Indig F.E., Jozwiak K., Bernier M, & Wainer I.W. (2017). Concurrent activation of β2-adrenergic receptor and blockage of GPR55 disrupts pro-oncogenic signaling in glioma cells. Cellular signalling, 36, 176-188.

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10

Modulating Descending Serotonergic Pathways in Pain

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Desipramine (Sigma D3900; i.p. 25 mg/kg per 0.9% saline), a norepinephrine reuptake inhibitor, was administered 1 hour before 5,7-dihydroxytryptamine creatinine sulphate (5,7-DHT) to prevent toxicity to noradrenergic neurons. 5,7-DHT (Sigma 37970; i.t. 60 μg in 10 μL/0.9% saline) was used to ablate descending serotonergic fibres from the rostral ventromedial medulla (RVM) and administered 7 days before formalin stimulation. SL327 (Tocris 1969; i.p. 50 mg/kg per DMSO; Tocris Bioscience, Bristol, United Kingdom) and SB747651A (Tocris 4630; i.t. 10 μM in 10 μL or 1 μM in 10 μM per 0.0086% ethanol per 0.9% saline) were delivered 30 minutes before formalin stimulation.

Tochiki K.K., Maiarú M., Norris C., Hunt S.P, & Géranton S.M. (2016). The mitogen and stress-activated protein kinase 1 regulates the rapid epigenetic tagging of dorsal horn neurons and nocifensive behaviour. Pain, 157(11), 2594-2604.

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Sl327

Lab products found in correlation

17 protocols using sl327

PTZ-Induced Seizure Assay in Mice

Preparation of Amyloid-β Peptide and Kinase Inhibitors

Ethanol and Inhibitor Preparation

Neuromodulatory Drugs in Behavioral Assays

Neuromodulatory Drugs in Behavioral Assays

Pharmacological Evaluation of Spatial Learning

Evaluating MEK1/2 Inhibitor Neuroprotection

Modulation of Neuronal Activity

Synthesis of Enantiomeric Adrenergic Compounds

Modulating Descending Serotonergic Pathways in Pain

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