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Thiamet g

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Thiamet-G is a laboratory reagent produced by Bio-Techne. It is an inhibitor of the O-GlcNAcase enzyme, which is responsible for the removal of O-linked N-acetylglucosamine (O-GlcNAc) modifications from proteins. Thiamet-G can be used in research applications to study the role of O-GlcNAc signaling in various biological processes.

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Lab products found in correlation

Perchloric acid, by Thermo Fisher Scientific (2 mentions) Medetomidine, by Henry Schein (2 mentions) Sodium dodecyl sulfate (sds), by Teknova (2 mentions) Sodium deoxycholate, by Merck Group (2 mentions) Methanol, by Thermo Fisher Scientific (2 mentions) Trichostatin a, by Merck Group (2 mentions) Halt protease and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (2 mentions) 4 2 hydroxyethyl piperazine 1 ethanesulfonic acid hepes, by Merck Group (2 mentions) Asp n, by Roche (2 mentions) Phosphatase inhibitor cocktails 1 or 3, by Merck Group (2 mentions) Dimethylamino phenyl n hydroxy 4 6 dimethyl 7 oxo 2 4 heptadienamide, by Merck Group (2 mentions) Niacinamide pyridine 3 carboxylic acid amide, by Merck Group (2 mentions) Proteomic grade trypsin, by Promega (2 mentions) Triton x 100, by Merck Group (2 mentions) Complete mini, by Roche (2 mentions) Pugnac, by Merck Group (2 mentions) Toluidine blue, by Merck Group (1 mentions) Osmi 1, by Merck Group (1 mentions) Eclipse 80i microscope, by Nikon (1 mentions) Bone slices, by Immunodiagnostic Systems (1 mentions)

6 protocols using thiamet g

1

In vitro Osteoclastogenesis Assay

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The bone resorption assays were performed as previously described75 (link). In brief, bone marrow cells from mice were seeded on bone slices (Immunodiagnostic Systems, Frankfurt am Main, Germany) with cytokines for in vitro osteoclastogenesis and with or without 20 µmol·L−1 OSMI-1 (Sigma-Aldrich) or 10 µmol·L−1 Thiamet-G (Tocris, Wiesbaden-Nordenstadt, Germany). The culture medium was changed every 2 days for 2 weeks. The resorption pits were visualized by staining with 1% toluidine blue (Sigma-Aldrich). Images were acquired using a Nikon Eclipse 80i microscope (Nikon Metrology) and then analyzed with ImageJ software (NIH, version 1.52p).
Li Y.N., Chen C.W., Trinh-Minh T., Zhu H., Matei A.E., Györfi A.H., Kuwert F., Hubel P., Ding X., Manh C.T., Xu X., Liebel C., Fedorchenko V., Liang R., Huang K., Pfannstiel J., Huang M.C., Lin N.Y., Ramming A., Schett G, & Distler J.H. (2022). Dynamic changes in O-GlcNAcylation regulate osteoclast differentiation and bone loss via nucleoporin 153. Bone Research, 10, 51.
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2

Protein Extraction and Modification Assay

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Phosphatase inhibitor cocktails 1 or 3, and 2 (100x, Sigma) and protease inhibitor tablets (cOmplete, EDTA-free and cOmplete-mini, EDTA-free; Roche) were used interchangeably with Halt protease and phosphatase inhibitor cocktails (Thermo Scientific); all were used at 1x according to the manufacturers’ instructions. Thiamet G (4390, Tocris) was used in a range of concentrations, as specified. Trichostatin A ([R-(E,E)]-7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxo-2,4-heptadienamide, Sigma) was used at 3 μM and niacinamide (pyridine-3-carboxylic acid amide, Sigma) at 10 mM. Other reagents were ketamine HCl (Bionichepharma), medetomidine (Dexdomitor, Henry Schein), perchloric acid (Fisher), methanol (Fisher), 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES, Sigma), sodium chloride (5M, Cellgrow), sodium dodecyl sulfate (from 10% solution, Teknova), TritonX-100 (Sigma), and sodium deoxycholate (Sigma). All reagents for peptide digestions were from Sigma, except proteomic-grade trypsin (Promega) and AspN (Roche).
Morris M., Knudsen G.M., Maeda S., Trinidad J.C., Ioanoviciu A., Burlingame A.L, & Mucke L. (2015). Tau post-translational modifications in wildtype and human amyloid precursor protein transgenic mice. Nature neuroscience, 18(8), 1183-1189.
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3

Glycosidase Inhibitors in Biological Study

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PUGNAc (Sigma-Aldrich; A7229), OSMI-1 (GLIXX Lab; Southborough, MA; GLXC-06077) and Thiamet-G (TOCRIS; Bristol, UK; 4390) were used in the study.
Das S., Bailey S.K., Metge B.J., Hanna A., Hinshaw D.C., Mota M., Forero-Torres A., Chatham J.C., Samant R.S, & Shevde L.A. (2018). O-GlcNAcylation of GLI Transcription Factors in Hyperglycemic Conditions Augments Hedgehog activity: Hyperglycemic conditions upregulate GLI activity. Laboratory investigation; a journal of technical methods and pathology, 99(2), 260-270.
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4

Glycosidase Inhibitors in Biological Study

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PUGNAc (Sigma-Aldrich; A7229), OSMI-1 (GLIXX Lab; Southborough, MA; GLXC-06077) and Thiamet-G (TOCRIS; Bristol, UK; 4390) were used in the study.
Das S., Bailey S.K., Metge B.J., Hanna A., Hinshaw D.C., Mota M., Forero-Torres A., Chatham J.C., Samant R.S, & Shevde L.A. (2018). O-GlcNAcylation of GLI Transcription Factors in Hyperglycemic Conditions Augments Hedgehog activity: Hyperglycemic conditions upregulate GLI activity. Laboratory investigation; a journal of technical methods and pathology, 99(2), 260-270.
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5

Investigating TRPM7 and O-GlcNAcylation

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2-APB, SKF96365, and thiamet G were obtained from Tocris Bioscience (Bristol, UK). PugNAc and antibodies for TRPM7, ORAI1, STIM1, OGA, O-GlcNAc and ITGB7 were obtained from Abcam (Cambridge, UK). Antibody for ubiquitin was from Santa Cruz Biotechnology (Dallas, TX, USA) and secondary antibodies were from EMD Millipore (Berlington, MA, USA). MG-132 and all other antibodies were from Cell Signaling Technology (Beverly, MA, USA). Other reagents were from Sigma-Aldrich (Dallas, MA, USA).
Samart P., Luanpitpong S., Rojanasakul Y, & Issaragrisil S. (2021). O-GlcNAcylation homeostasis controlled by calcium influx channels regulates multiple myeloma dissemination. Journal of Experimental & Clinical Cancer Research : CR, 40, 100.
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6

Protein Extraction and Modification Assay

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Phosphatase inhibitor cocktails 1 or 3, and 2 (100x, Sigma) and protease inhibitor tablets (cOmplete, EDTA-free and cOmplete-mini, EDTA-free; Roche) were used interchangeably with Halt protease and phosphatase inhibitor cocktails (Thermo Scientific); all were used at 1x according to the manufacturers’ instructions. Thiamet G (4390, Tocris) was used in a range of concentrations, as specified. Trichostatin A ([R-(E,E)]-7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxo-2,4-heptadienamide, Sigma) was used at 3 μM and niacinamide (pyridine-3-carboxylic acid amide, Sigma) at 10 mM. Other reagents were ketamine HCl (Bionichepharma), medetomidine (Dexdomitor, Henry Schein), perchloric acid (Fisher), methanol (Fisher), 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES, Sigma), sodium chloride (5M, Cellgrow), sodium dodecyl sulfate (from 10% solution, Teknova), TritonX-100 (Sigma), and sodium deoxycholate (Sigma). All reagents for peptide digestions were from Sigma, except proteomic-grade trypsin (Promega) and AspN (Roche).
Morris M., Knudsen G.M., Maeda S., Trinidad J.C., Ioanoviciu A., Burlingame A.L, & Mucke L. (2015). Tau post-translational modifications in wildtype and human amyloid precursor protein transgenic mice. Nature neuroscience, 18(8), 1183-1189.
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