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Bay 559837

Manufactured by Bio-Techne
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Sourced in United Kingdom

BAY-559837 is a laboratory instrument manufactured by Bio-Techne. The core function of this product is to perform analytical measurements and data processing for scientific research applications.

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Lab products found in correlation

Aria 2 facs sorter, by BD (2 mentions) Cytofix cytoperm buffer set, by BD (2 mentions) β mercaptoethanol, by Thermo Fisher Scientific (2 mentions) Ionomycin, by Merck Group (2 mentions) Golgiplug, by BD (2 mentions) Il 23, by R&D Systems (2 mentions) Sp600125, by Bio-Techne (2 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions) Brain derived neurotrophic factor (bdnf), by R&D Systems (1 mentions) Sch772984, by Cayman Chemical (1 mentions) Tetrodotoxin ttx, by Merck Group (1 mentions) Rp 8 br camps, by Biolog (1 mentions) Glutamate, by Bio-Techne (1 mentions) Myristoylated pki 14 22 amide, by Bio-Techne (1 mentions) Sotrastaurin, by Cayman Chemical (1 mentions) Sodium hydroxide (naoh), by Merck Group (1 mentions) Total p38, by Cell Signaling Technology (1 mentions) Phospho p38, by Cell Signaling Technology (1 mentions) Sb239063, by Merck Group (1 mentions) Tubulin, by Abcam (1 mentions)

6 protocols using bay 559837

1

Isolation and Culture of Murine ILC3

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CCR6+ and CCR6neg ILC3 (DAPInegCD3negCD11cnegCD14negCD19negTCRβnegTCRγnegNK1.1negKLRG1negCD127+CD90.2+) were isolated from small intestine LPLs of C57BL/6 mice using the ARIA II FACS Sorter (BD Biosciences). ILC3 were cultured at 37°C in flat bottom 96 well plates (104 cells/well) in RPMI supplemented with 10% heat-inactivated FBS (Hyclone), 50 U penicillin-streptomycin (Hyclone), 2 mM glutamine (Hyclone), 10mM HEPES (Hyclone), 1mM sodium pyruvate (Hyclone) and 50 μM β-mercaptoethanol (Gibco). ILC3 were stimulated with IL-23 (100–300 pg/mL, R&D systems) and/or VIPR2 ligands (BAY-559837: 1–100nM, and VIP: 1nM-1uM, TOCRIS) for 16h (37°C), washed, incubated in for 4h at 37°C in RPMI with 10% FBS, phorbol 12-myristate 13-acetate (PMA) (50 ng/ml; Sigma), ionomycin (500 ng/ml;Sigma) and Golgi Plug (BD Bioscience), and stained for membrane extracellular markers in Staining Buffer (PBS FBS 2% EDTA 5mM) and for intracellular markers using Cytofix/Cytoperm buffer set following manufacturer’s protocol (BD Biosciences). Acquisition of cytometric parameters was performed on an LSRII (BD Biosciences). All data were analyzed using FlowJo Software Version 10 (Tree Star).
Talbot J., Hahn P., Kroehling L., Nguyen H., Li D, & Littman D.R. (2020). Feeding-dependent VIP neuron-ILC3 circuit regulates the intestinal barrier. Nature, 579(7800), 575-580.
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2

Pharmacological Modulation of Circadian Rhythms

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VIP (Tocris) and Bay 55-9837 (Tocris) stocks were prepared by dissolving in HEPES-buffered medium. Glutamate (1 mM; Tocris) was dissolved in 1eq. NaOH, Tetrodotoxin (TTX, 1 µM; Sigma) and BDNF (200 ng/ml; R&D Systems) were dissolved in water. BDNF was applied at CT16, while TTX was applied 24 h prior to VIP treatment. Sotrastaurin (300 nM; Cayman Chemical), SP600125 (3 µM; Tocris), SCH772984 (100 nM; Cayman Chemical) and 666-15 (1 µM; Tocris) were all dissolved in DMSO, Rp-8-Br-cAMPS (50 µM; Biolog) in water, while PKI 14-22 amide myristoylated (1 µM; Tocris) was dissolved in 30% acetonitrile; these 6 drugs were used 30 min prior to VIP treatment. All pharmacological agents were bath-applied to SCN slices unless otherwise stated, and washed off only if stated explicitly. Phase of treatments was extrapolated from the preceding rhythm. Due to their arrhythmic nature, Cry1−/−Cry2−/− slices were treated based on Cry1−/−Cry2+/- littermate phases (data not shown).
Hamnett R., Crosby P., Chesham J.E, & Hastings M.H. (2019). Vasoactive intestinal peptide controls the suprachiasmatic circadian clock network via ERK1/2 and DUSP4 signalling. Nature Communications, 10, 542.
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3

Evaluation of p38 Inhibitor Efficacy

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BAY 55-9837 was purchased from Tocris Bioscience. p38 inhibitor SB239063 was purchased from Sigma. The antibodies used in this study were SMN/Smn (BD Transduction Laboratories), Actin (Abcam), Tubulin (Abcam), Phospho-p38 (Cell signalling) and Total p38 (Cell signalling).
Hadwen J., MacKenzie D., Shamim F., Mongeon K., Holcik M., MacKenzie A, & Farooq F. (2014). VPAC2 receptor agonist BAY 55-9837 increases SMN protein levels and moderates disease phenotype in severe spinal muscular atrophy mouse models. Orphanet Journal of Rare Diseases, 9, 4.
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4

Macrophage Polarization by VIP Signaling

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RAW264.7 cells (2 × 104) were incubated with 20% conditioned medium derived from CT26 with DMEM complete medium (CT26-CM) or 20% RPMI medium with DMEM complete medium (control). VIP hybrid (KPRRPYTDNYTRLRKQMAVKKYLNSILN-amide, Phoenix Pharmaceutical, Burlingame, CA)—a competitive antagonist of VIP17 (link)—at 1 and 3 µM was added daily. In a separate experiment, [Lys15, Arg16, Leu27] VIP (1–7)/GRF (8–27) (Phoenix Pharmaceuticals)—a selective VPAC1 agonist30 (link)—or BAY 55-9837 (Tocris Bioscience, Minneapolis, MN)—a selective VPAC2 agonist—was added daily at a dose of 100 nM31 (link). mRNA extraction was performed on day 3 or day 4 after CT26-CM incubation, depending on the experiment. PBS was used as vehicle treatment.
Vipr1, Vipr2 or negative control siRNA (Thermo Fisher Scientific) was transfected into RAW264.7 cells at day 1 after CT26-CM incubation using Lipofectamine RNAimax (Thermo Fisher Scientific) following the manufacturer’s instructions. Vipr1 or Vipr2 mRNA expression was analyzed to evaluate knock-down efficiency at day 3 after CT26-CM incubation. M1 and M2 gene expression was measured at day 4 after CT26-CM incubation.
Kittikulsuth W., Nakano D., Kitada K., Uyama T., Ueda N., Asano E., Okano K., Matsuda Y, & Nishiyama A. (2023). Vasoactive intestinal peptide blockade suppresses tumor growth by regulating macrophage polarization and function in CT26 tumor-bearing mice. Scientific Reports, 13, 927.
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5

Isolation and Culture of Murine ILC3

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CCR6+ and CCR6neg ILC3 (DAPInegCD3negCD11cnegCD14negCD19negTCRβnegTCRγnegNK1.1negKLRG1negCD127+CD90.2+) were isolated from small intestine LPLs of C57BL/6 mice using the ARIA II FACS Sorter (BD Biosciences). ILC3 were cultured at 37°C in flat bottom 96 well plates (104 cells/well) in RPMI supplemented with 10% heat-inactivated FBS (Hyclone), 50 U penicillin-streptomycin (Hyclone), 2 mM glutamine (Hyclone), 10mM HEPES (Hyclone), 1mM sodium pyruvate (Hyclone) and 50 μM β-mercaptoethanol (Gibco). ILC3 were stimulated with IL-23 (100–300 pg/mL, R&D systems) and/or VIPR2 ligands (BAY-559837: 1–100nM, and VIP: 1nM-1uM, TOCRIS) for 16h (37°C), washed, incubated in for 4h at 37°C in RPMI with 10% FBS, phorbol 12-myristate 13-acetate (PMA) (50 ng/ml; Sigma), ionomycin (500 ng/ml;Sigma) and Golgi Plug (BD Bioscience), and stained for membrane extracellular markers in Staining Buffer (PBS FBS 2% EDTA 5mM) and for intracellular markers using Cytofix/Cytoperm buffer set following manufacturer’s protocol (BD Biosciences). Acquisition of cytometric parameters was performed on an LSRII (BD Biosciences). All data were analyzed using FlowJo Software Version 10 (Tree Star).
Talbot J., Hahn P., Kroehling L., Nguyen H., Li D, & Littman D.R. (2020). Feeding-dependent VIP neuron-ILC3 circuit regulates the intestinal barrier. Nature, 579(7800), 575-580.
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6

Inhibition Kinetics of Tissue Transglutaminase

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Akt inhibitor XI, KT 5720, were purchased from Calbiochem (San Diego, CA). [Ala 11,22,28 ]-VIP, Bay 55-9837, PACAP-27, PACAP 6-38, PD 98059, SB 203580, and SP 600 125 were obtained from Tocris Bioscience (Bristol, UK). All-trans retinoic acid, casein, Protease Inhibitor Cocktail (for use with mammalian cell and tissue extracts), Phosphatase Inhibitor Cocktail 2 and 3, ExtrAvidin ® -HRP and ExtrAvidin ® -FITC were obtained from Sigma-Aldrich Co. Ltd. (Gillingham, UK). The TG2 inhibitors Z-DON (Z-ZON-Val-Pro-Leu-OMe) and R283, together with purified guinea-pig liver TG2 were obtained from Zedira GmbH (Darmstadt, Germany). DAPI was from Vector Laboratories Inc (Peterborough, UK). Fluo-8/AM was purchased from Stratech Scientific Ltd (Newmarket, UK). Biotin-TVQQEL was purchased from Pepceuticals (Enderby, UK). Biotin Biotechnology Inc (Heidelberg, Germany).
Algarni A.S., Hargreaves A.J, & Dickenson J.M. (2017). Role of transglutaminase 2 in PAC1 receptor mediated protection against hypoxia-induced cell death and neurite outgrowth in differentiating N2a neuroblastoma cells. Biochemical pharmacology, 128.
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