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5 iodotubercidin

Manufactured by Merck Group
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Sourced in Switzerland, United States

5-iodotubercidin is a synthetic compound used as a laboratory reagent. It is a nucleoside analogue that can inhibit the activity of certain enzymes. The core function of 5-iodotubercidin is to serve as a research tool for biochemical and cell biology studies. Further details on its intended use are not provided.

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Lab products found in correlation

Dipyridamole, by Merck Group (2 mentions) Adenosine triphosphate (atp), by Merck Group (1 mentions) Adenosine, by Merck Group (1 mentions) Adenosine diphosphate (adp), by Merck Group (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Rpmi 1640 with l glutamine, by Thermo Fisher Scientific (1 mentions) Negative selection columns for neutrophil isolation, by STEMCELL (1 mentions) 5z 7 oxozeaenol, by Merck Group (1 mentions) Aicar, by Enzo Life Sciences (1 mentions) Cgs 15943, by Merck Group (1 mentions) Colchicine, by Merck Group (1 mentions) Nocodazole, by Merck Group (1 mentions) Podophyllotoxin, by Merck Group (1 mentions) Tnf α, by Thermo Fisher Scientific (1 mentions) Screen well kinase inhibitor library, by Enzo Life Sciences (1 mentions) Gsk983, by Merck Group (1 mentions) Tert butylhydroquinone tbhq, by Merck Group (1 mentions)

4 protocols using 5 iodotubercidin

1

Molecular Pharmacology of Adenosine Receptors

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Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) was purchased from Enzo Life Sciences (Lausen, Switzerland), whereas 5-iodotubercidin, dipyridamole, and standard compounds of high-performance liquid chromatography grade (adenosine, ATP, ADP, cAMP, AMP) were from Sigma-Aldrich (St. Louis, MO). The sources of other reagents and chemicals are listed in Malaga-Dieguez et al. (2010) (link), which also contains a description of the constructs encoding wild-type and mutated versions of the human A1-adenosine receptor fused to green fluorescent protein GFP. In addition, we generated N-terminally modified versions by addition of either a FLAG epitope or a Strep-Tactin II-FLAG tandem affinity purification tag (SF-TAP) at the N terminus (Gloeckner et al., 2007 (link)). Mutant V2R constructs were a generous gift from Ralf Schülein (Leibniz Institute for Molecular Pharmacology, Berlin, Germany). An N-terminal FLAG tag was added to these receptor constructs by subcloning their cDNA into the Tag-2B expression vector (Stratagene, La Jolla, CA). The integrity of the DNA constructs was verified by fluorescent sequencing.
Kusek J., Yang Q., Witek M., Gruber C.W., Nanoff C, & Freissmuth M. (2014). Chaperoning of the A1-Adenosine Receptor by Endogenous Adenosine—An Extension of the Retaliatory Metabolite Concept. Molecular pharmacology, 87(1), 39-51.
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2

AICAR-Mediated AMPK and p38 Activation

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AICAR was purchased from Enzo Life Sciences (Plymouth Meeting, PA, USA). RPMI 1640 with L-glutamine, DMEM, penicillin-streptomycin and fetal bovine serum (FBS) were obtained from GIBCO (Gaithersburg, MD, USA). Bicinchoninic acid (BCA) protein assay reagent was obtained from Pierce (Rockford, IL, USA). Antibodies specific for p-AMPK (Thr172), p-p38 (Thr180/Tyr182), p-MKK3/6 (Ser189/Ser207), AMPK and p38, and compound C were purchased from Cell Signaling Technologies (Beverly, MA, USA). CGS15943, 5’-iodotubercidin, (5z)-7-oxozeaenol and dipyridamole were from Sigma-Aldrich (St Louis, MO, USA). SB203580 was obtained from Calbiochem (La Jolla, CA, USA). Control siRNA and siRNA to the AMPKα1 and p38α were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Custom antibody mixtures and negative selection columns for neutrophil isolation were purchased from Stem cell Technologies (Vancouver, British Columbia, Canada).
Quan H., Kim J.M., Lee H.J., Lee S.H., Choi J.I, & Bae H.B. (2015). AICAR Enhances the Phagocytic Ability of Macrophages towards Apoptotic Cells through P38 Mitogen Activated Protein Kinase Activation Independent of AMP-Activated Protein Kinase. PLoS ONE, 10(5), e0127885.
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3

Quantifying CD73 Activity in Tumors

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Implanted B16F10 tumors were excised, snap frozen, and homogenized in protein lysis buffer. Inhibitors including 10 μmol/L EHNA, 625 μmol/L TNAP inhibitor (supplied by Arcus Biosciences), 10 μmol/L 5-iodotubercidin (Sigma-Aldrich), and 4 μmol/L aristeromycin (Sigma-Aldrich) were added to the lysate at 37°C, pH 7.4. AB680 was added exogenously to tumor homogenates and incubated for 1 hour at 37°C before adding 5 μmol/L 13C5 AMP. After 5 minutes, the reaction was quenched with a final concentration of 0.1 mol/L PCA and CD73 activity was calculated from the amount of 13C5 adenosine produced (measured by LC/MS).
Piovesan D., Tan J.B., Becker A., Banuelos J., Narasappa N., DiRenzo D., Zhang K., Chen A., Ginn E., Udyavar A.R., Yin F., Paprcka S.L., Purandare B., Park T.W., Kimura N., Kalisiak J., Young S.W., Powers J.P., Schindler U., Sivick K.E, & Walters M.J. (2022). Targeting CD73 with AB680 (Quemliclustat), a Novel and Potent Small-Molecule CD73 Inhibitor, Restores Immune Functionality and Facilitates Antitumor Immunity. Molecular Cancer Therapeutics, 21(6), 948-959.
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4

High-Throughput Screening of Kinase Inhibitors

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The chemical libraries used for the screens were the SCREEN-WELL Kinase Inhibitor library (Enzo Life Sciences), the Prestwick Chemical library (Prestwick Chemical), and a subset of Microbiotix libraries of >200,000 discrete compounds previously used in high-throughput screening for inhibitors of anti-infective targets33 (link)–39 (link). GSK983, podophyllotoxin, colchicine, nocodazole, 5-iodotubercidin, tert-butylhydroquinone (tBHQ), uridine, deoxycytidine and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich. F0015–0327, F2336–0041, F5540–0057, F5001–2941, C798–0517, E975–1448, F2468–0184, F2468–0196, 6259503, F5139–0048, F5139–0096, J107–0137, J107–0140, J107–0181 and J107–0307 were purchased from Life Chemicals Inc. All compounds were diluted to a concentration of 20 mM in DMSO before use. tBHQ was diluted in ethanol to a concentration of 50 mM. Uridine and deoxycytidine were diluted to 10 mM in distilled water. TNFα was purchased from PeproTech.
Edwards M.R., Liu G., De S., Sourimant J., Pietzsch C., Johnson B., Amarasinghe G.K., Leung D.W., Bukreyev A., Plemper R.K., Aron Z., Bowlin T.L., Moir D.T, & Basler C.F. (2020). Small molecule compounds that inhibit antioxidant response gene expression in an inducer-dependent manner. ACS infectious diseases, 6(3), 489-502.
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