4-Thiouridine (Synonyms: NSC 518132, 4-SU) |
Katalog-Nr.GC42474
One-Click Copy Product Info
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4-Thiouridin ist ein Ribonukleosid-Analogon, es wird hÄufig in der RNA-Analyse und (m)RNA-Markierung verwendet.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 13957-31-8
Sample solution is provided at 25 µL, 10mM.
4-Thiouridine(4sU) is a ribonucleoside analogue that can be used for RNA analysis and (m)RNA labeling[1]. 4-Thiouridine is a sulfur-containing uridine derivative in which the oxygen atom at the 4-position of the uridine ring is replaced with a sulfur atom. This modification is naturally found in certain RNA molecules, particularly in bacterial transfer RNA (tRNA), where it plays a role in stabilizing RNA structure and enhancing its function[2-3].
In laboratory applications, 4-thiouridine serves as a labeling tool to study RNA synthesis, metabolism, and localization[4-5]. Upon UV irradiation, s4U-labeled RNA can form covalent cross-links with adjacent molecules, such as proteins, which facilitates the analysis of RNA interactions and dynamics within cells[6].
4-Thiouridine can interfere with nucleolar integrity and trigger a nucleolar stress response. 4-Thiouridine triggers translocation of NPM1, stabilization of p53, and inhibition of proliferation [7].
References:
[1]. Dölken L, Ruzsics Z, et,al. High-resolution gene expression profiling for simultaneous kinetic parameter analysis of RNA synthesis and decay. RNA. 2008 Sep;14(9):1959-72. doi: 10.1261/rna.1136108. Epub 2008 Jul 24. PMID: 18658122; PMCID: PMC2525961.
[2]. Hafner M, Landthaler M, et,al. Transcriptome-wide identification of RNA-binding protein and microRNA target sites by PAR-CLIP. Cell. 2010 Apr 2;141(1):129-41. doi: 10.1016/j.cell.2010.03.009. PMID: 20371350; PMCID: PMC2861495.
[3]. Windhager L, Bonfert T, et,al. Ultrashort and progressive 4sU-tagging reveals key characteristics of RNA processing at nucleotide resolution. Genome Res. 2012 Oct;22(10):2031-42. doi: 10.1101/gr.131847.111. Epub 2012 Apr 26. PMID: 22539649; PMCID: PMC3460197.
[4]. Rädle B, Rutkowski AJ, et,al. Metabolic labeling of newly transcribed RNA for high resolution gene expression profiling of RNA synthesis, processing and decay in cell culture. J Vis Exp. 2013 Aug 8;(78):50195. doi: 10.3791/50195. PMID: 23963265; PMCID: PMC3854562.
[5]. Schwanhäusser B, Busse D, et,al. Global quantification of mammalian gene expression control. Nature. 2011 May 19;473(7347):337-42. doi: 10.1038/nature10098. Erratum in: Nature. 2013 Mar 07;495(7439):126-7. doi: 10.1038/nature11848. PMID: 21593866.
[6]. Sun W, Chen W. Metabolic Labeling of Newly Synthesized RNA with 4sU to in Parallel Assess RNA Transcription and Decay. Methods Mol Biol. 2018;1720:25-34. doi: 10.1007/978-1-4939-7540-2_3. PMID: 29236249.
[7]. Burger K, Mühl B, et,al. 4-thiouridine inhibits rRNA synthesis and causes a nucleolar stress response. RNA Biol. 2013 Oct;10(10):1623-30. doi: 10.4161/rna.26214. Epub 2013 Sep 4. PMID: 24025460; PMCID: PMC3866244.
Ⅰ. Protocol for 4-thiouridine tagging [1]:
4sU solution: Use 4sU powder to prepare the solution as 50mM stock in ddH2O(double distilled water) and 0.25um filtration to remove bacteria. Keep in dark place. Store the solution at −20 ℃ in aliquots of 100μl.
1．1.5 × 106 2fTGH cells were cultured in presence of 4-thiouridine (10µM), lysed, and total RNA was extracted as described for metabolic in vivo labeling.
2．50μg total RNA was incubated with biotin-HPDP (1mg/ml; 2μl/μg RNA) in biotinylation buffer (100mM Tris, 10mM EDTA, pH 7.4, 1μl/μg RNA) for 1.5h at RT.
3．An equal volume of chloroform was added, mixed, and incubated with biotinylated RNA for 3min. The mixture was separated in pre-spun Phase Trap Gel Heavy Tubes (5min, 16000rpm).
4．For RNA precipitation and removal of unincorporated biotin-HPDP, a 1/10 volume 5M NaCl and an equal volume of absolute isopropanol were added to the aqueous phase and centrifuged (20min, 16000rpm).
5．The pellet was washed in an equal volume of 75% ethanol and centrifuged (10min, 16000rpm).
6．RNA was resuspended in 100μl RNase-free H2O.
7．For separation, untagged and 4-thiouridine-tagged RNA was first heated to 65℃ for 10min and cooled on ice for 5min. RNA was incubated with 75μl streptavidin-coated magnetic beads for 15min with rotation.
8．The reaction volume was applied to μMACs columns, placed in an OctoMACS Seperator magnetic stand, and equilibrated with 900μl μMACS washing buffer (100mM Tris, 10mMEDTA, 1M NaCl, 0.1% Tween-20, pH 7.5).
9．The columns were washed with μMACS washing buffer.
10. 4-thiouridine-biotin-streptavidin-tagged RNA was eluted in 700μl RLT lysis buffer with dithioerythritol (100mM).
11. 4-thiouridine-tagged RNA was recovered with the PeqGOLD total RNA kit.
12. 4-thiouridine-tagged RNA was separated on a 1.5% agarose-gel, containing ethidium bromide (37.5μg/100ml).
13. Signals of RNA under UV-light were quantified by AIDA software.
This program only provides a guide, please modify according to your specific needs.
References:
[1]. Burger K, Mühl B, et,al. 4-thiouridine inhibits rRNA synthesis and causes a nucleolar stress response. RNA Biol. 2013 Oct;10(10):1623-30. doi: 10.4161/rna.26214. Epub 2013 Sep 4. PMID: 24025460; PMCID: PMC3866244.
| Cas No. | 13957-31-8 | SDF | |
| Überlieferungen | NSC 518132, 4-SU | ||
| Canonical SMILES | OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C=CC(NC2=O)=S)O1 | ||
| Formula | C9H12N2O5S | M.Wt | 260.3 |
| Löslichkeit | DMF: 10 mg/ml,DMSO: 10 mg/ml,Ethanol: 2 mg/ml,PBS (pH 7.2): 5 mg/ml | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 3.8417 mL | 19.2086 mL | 38.4172 mL |
| 5 mM | 768.3 μL | 3.8417 mL | 7.6834 mL |
| 10 mM | 384.2 μL | 1.9209 mL | 3.8417 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
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Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
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- Purity: >98.00% Appearance: A solid
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- SDS (Safety Data Sheet)
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Average Rating: 5 (Based on Reviews and 1 reference(s) in Google Scholar.)