Mouse anti-human IFNγ IgG1 and IgG1 isotype control were from Biolegend. 5′-Deoxy-5′-(methylthio)adenosine (MTA) and rapamycin were from Sigma.
5 deoxy 5 methylthio adenosine mta
Manufactured by Merck Group
Sourced in United States, China
5′-Deoxy-5′-(methylthio)adenosine (MTA) is a chemical compound used as a research tool in various scientific applications. It is a naturally occurring sulfur-containing adenosine derivative. MTA serves as a core component in biochemical and cell biology studies, but its specific intended use should not be extrapolated upon.
Automatically generated - may contain errors
Lab products found in correlation
Igg1 isotype control, by BioLegend (1 mentions)
Rapamycin, by Merck Group (1 mentions)
3 deazaneplanocin a dznep, by Cayman Chemical (1 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions)
5 aza 2 deoxycytidine, by Merck Group (1 mentions)
Inosine, by Merck Group (1 mentions)
Abt 702, by Bio-Techne (1 mentions)
Mrs1754, by Bio-Techne (1 mentions)
Nitrobenzylthioinosine nbmpr, by Bio-Techne (1 mentions)
Adenosine 2 3 dialdehyde adox, by Merck Group (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Thp 1 cell, by American Type Culture Collection (1 mentions)
Huvecs, by American Type Culture Collection (1 mentions)
Adenosine, by Merck Group (1 mentions)
Cgs21680, by Bio-Techne (1 mentions)
Egm 2, by Lonza (1 mentions)
Tnf α, by R&D Systems (1 mentions)
Zm241385, by Bio-Techne (1 mentions)
Il 1β, by R&D Systems (1 mentions)
Trichostatin a tsa, by Beyotime (1 mentions)
5 protocols using 5 deoxy 5 methylthio adenosine mta
1
Evaluating Interferon-gamma Signaling
2
Epigenetic Chromatin Remodeling in Stem Cells
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hMSCs and HFF-1s were treated with lysine methyltransferase inhibitors (KMTi) to forcibly assume a chromatin structural state that is either more open or more closed. Cells were treated with 1, 5 and 10 μM of 3-Deazaneplanocin A (DZNep) (Cayman Chemical, Ann Arbor, Michigan, USA) for 72 hours to selectively inhibit the enzymatic activity of EZH2, which is a KMT that targets lysine residue 27 on histone 3. 1, 2 and 4 mM of 5′-Deoxy-5′-(methylthio)adenosine (MTA) (Sigma-Aldrich) was introduced to cells for 24 hours to inhibit the MLL family of enzymes, which catalyze the trimethylation of lysine residue 4 on histone 3. After drug treatment, cells were subsequently fixed in 4% Paraformaldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) and prepared for immunocytochemical labeling.
3
Endothelial Cell Culture and Pharmacological Modulation
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HUVECs (ATCC, Manassas, VA, USA), at passage 3–8, and MAECs, at passage 2–4, were cultured in endothelial growth medium 2 (EGM-2; Lonza, Basel, Switzerland). In some experiments, 10 ng/ml TNF-α (R & D Systems, Minneapolis, MN, USA), 10 ng/ml IL-1β (R & D Systems, Minneapolis, MN, USA), 0–100 µM adenosine (Sigma, St Louis, MO, USA), 10 μM ITU (Tocris Bioscience, Bristol, United Kingdom), 2 µM ABT702 (Tocris Bioscience, Bristol, United Kingdom), 5 μM CGS21680 (Tocris Bioscience, Bristol, United Kingdom), 5 μM NECA (Tocris Bioscience, Bristol, United Kingdom), 5 μM MRS1754 (Tocris Bioscience, Bristol, United Kingdom), 5 μM ZM241385 (Tocris Bioscience, Bristol, United Kingdom), 10 μM nitrobenzylthioinosine (NBMPR) (Tocris Bioscience, Bristol, United Kingdom), 0.1–1mM inosine (Sigma, St Louis, MO, USA), 2 mM 5′-deoxy-5′ (methylthio)adenosine (MTA) (Sigma, St Louis, MO, USA), 2 μM 5-Aza-2′-deoxycytidine (Sigma, St Louis, MO, USA), 1 mM SAH (Sigma, St. Louis, MO, USA) or 20 µM adenosine-2′,3′-dialdehyde (Adox, Sigma, St Louis, MO, USA) was added to the culture medium. Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) used for monocyte adhesion assay were cultured in RPMI medium 1640 (Thermo Scientific, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS; Thermo Scientific, Grand Island, NY, USA).
4
Wound Healing Assay for Tenocyte Migration
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The migration was assessed through a wound healing assay. The tenocytes were seeded at a density of 4 × 104 cells/well in a 24-well culture plate. After reaching 80–85% confluence, the tenocytes were synchronized by serum starvation for 12 h. The confluent monolayer was then scratched with a 10-μL pipet tip and washed twice with PBS. The wells were filled with 1 mL of serum-free DMEM with or without MGF-C25E (Phoenix Pharmaceuticals, Burlingame, USA). To test whether chromatin decompaction inhibited the rate of cell migration, we examined the effect of the levels of DNA methylation on the rate of cell migration. The cells were incubated with histone deacetylase (HDAC) inhibitor trichostatin A (TSA) (Beyotime, Jiangsu, China) or methylase inhibitor 5’-deoxy-5’-methylthioadenosine (MTA) (Sigma-Aldrich, St. Louis, MO, USA) for 60 min at 37 °C before scratching. Images of the wounds were acquired at 0, 24, and 48 h through a microscope (Olympus, Japan). Using Image J, the levels of wound closure were assessed by calculating the ratio of the closure area to the initial wound area as follows: × 100 [%], where Wn represents the percentage of wound closure, An represents the residual wound area at the metering time point (h), and A0 represents the initial wound area.
5
Synthesis and Purification of MT-DADMe-ImmA
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MT-DADMe-ImmA was synthesized
as previously
described and generously provided by Dr. Gary B. Evans of the Carbohydrate
Chemistry Team, Ferrier Institute, Victoria University of Wellington,
Lower Hutt, New Zealand. Xanthine oxidase (Grade III) and 5′-deoxy-5′-methylthioadenosine
(MTA) were purchased from Sigma-Aldrich (Saint Louis, MO). 5,5′-Dithiobis-2-nitrobenzoic
acid (DTNB) was purchased from Sigma-Aldrich (Milwaukee, WI). All
other chemicals were purchased at the highest purity commercially
available and used without further purification.
as previously
described and generously provided by Dr. Gary B. Evans of the Carbohydrate
Chemistry Team, Ferrier Institute, Victoria University of Wellington,
Lower Hutt, New Zealand. Xanthine oxidase (Grade III) and 5′-deoxy-5′-methylthioadenosine
(MTA) were purchased from Sigma-Aldrich (Saint Louis, MO). 5,5′-Dithiobis-2-nitrobenzoic
acid (DTNB) was purchased from Sigma-Aldrich (Milwaukee, WI). All
other chemicals were purchased at the highest purity commercially
available and used without further purification.
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