| Size | Price | Stock | Qty |
|---|---|---|---|
| 500mg |
|
||
| Other Sizes |
|
Purity: ≥98%
| Targets |
Nucleoside Antimetabolite; DNA synthesis
|
|---|---|
| ln Vitro |
Replicating cells undergo DNA synthesis in the highly regulated, S-phase of the cell cycle. Analogues of the pyrimidine deoxynucleoside thymidine may be inserted into replicating DNA, effectively tagging dividing cells allowing their characterisation. Tritiated thymidine, targeted using autoradiography was technically demanding and superseded by 5-bromo-2-deoxyuridine (BrdU) and related halogenated analogues, detected using antibodies. Their detection required the denaturation of DNA, often constraining the outcome of investigations. Despite these limitations BrdU alone has been used to target newly synthesised DNA in over 20,000 reviewed biomedical studies. A recent breakthrough in "tagging DNA synthesis" is the thymidine analogue 5-ethynyl-2'-deoxyuridine (EdU). The alkyne group in EdU is readily detected using a fluorescent azide probe and copper catalysis using 'Huisgen's reaction' (1,3-dipolar cycloaddition or 'click chemistry'). This rapid, two-step biolabelling approach allows the tagging and imaging of DNA within cells whilst preserving the structural and molecular integrity of the cells. The bio-orthogonal detection of EdU allows its application in more experimental assays than previously possible with other "unnatural bases". These include physiological, anatomical and molecular biological experimentation in multiple fields including, stem cell research, cancer biology, and parasitology. The full potential of EdU and related molecules in biomedical research remains to be explored.[1]
|
| Cell Assay |
Tracking cell division through incorporating unnatural bases into newly synthesised DNA is a powerful and widely used tool in biology. Initial difficulties detecting tritiated thymidine was largely overcome using its halogenated analogues. The necessity for the immunodetection of these halogenated probes constrained studies, limiting them largely to anatomical investigations. The recent emergence of EdU, detected at high efficiency at near bio-orthogonal conditions, using selective “click chemistry”, allows investigation of DNA synthesis and cell genesis in a variety of assays. These have been extended to include living cells. These advances in tracking DNA synthesis are complemented by parallel advances in anatomical and molecular biology techniques. These include increasing sensitivity and automation of assays and detection technologies. Thus our capacity to investigate the most fundamental and secure process of life, DNA synthesis per-se is increased, facilitating the study of the replication and passage of life. The full potential of EdU and related molecules in biology and medicine, largely remain to be elucidated.
|
| References |
[1]. Cavanagh BL, et al. Thymidine analogues for tracking DNA synthesis. Molecules. 2011 Sep 15;16(9):7980-93.
|
| Additional Infomation |
Multicellular animals replicate their DNA when undergoing cell division. This occurs during embryogenesis and in the growth of tissues in the adult. DNA synthesis is the most primitive and defining event in the advent of the animal kingdom. The persistence of DNA over millions of years of animal evolution and its presence across all species alludes to its fundamental role and implies deep protective mechanisms to ensure its conservation. The four bases adenine, thymine, cytosine and guanine are the absolute unconditional coding components of DNA which is faithfully replicated during cell division, providing the robustly reproduced, blueprint for life.
DNA synthesis has a proof reading system that prevents the contamination of the bases. This tightly regulated process was initially breached using radiolabeled nucleoside analogs and subsequently with analogues detected using light microscopy. This allowed the tracking and subsequent characterization of newborn cells. During the S-phase of the cell cycle, the four DNA bases are organised by cellular machinery to form new DNA strands. The replicate paired strands are then segregated and drawn to the two poles of the mother cell prior to its cleavage to produce two daughter cells, each with a complement of newly synthesised DNA comprised of old and newly incorporated bases. If the cellular machinery is fooled into selecting and incorporating an “unnatural base” during DNA synthesis, it is possible to track the daughter cells and their offspring subsequent to that division.
|
| Molecular Formula |
C11H16N2O6
|
|---|---|
| Molecular Weight |
272.25
|
| Exact Mass |
272.1008
|
| Elemental Analysis |
C, 48.53; H, 5.92; N, 10.29; O, 35.26
|
| CAS # |
55486-09-4
|
| PubChem CID |
191372
|
| Appearance |
Typically exists as solids (or liquids in special cases) at room temperature
|
| Density |
1.5±0.1 g/cm3
|
| Index of Refraction |
1.597
|
| LogP |
-0.55
|
| tPSA |
113.78
|
| SMILES |
O1[C@]([H])(C([H])([H])O[H])[C@]([H])([C@]([H])([C@]1([H])N1C(N([H])C(C(C([H])([H])[H])=C1[H])=O)=O)OC([H])([H])[H])O[H]
|
| InChi Key |
YHRRPHCORALGKQ-FDDDBJFASA-N
|
| InChi Code |
InChI=1S/C11H16N2O6/c1-5-3-13(11(17)12-9(5)16)10-8(18-2)7(15)6(4-14)19-10/h3,6-8,10,14-15H,4H2,1-2H3,(H,12,16,17)/t6-,7-,8-,10-/m1/s1
|
| Chemical Name |
1-((2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-methoxytetrahydrofuran-2-yl)-5-methylpyrimidine-2,4(1H,3H)-dione
|
| Synonyms |
2'-O-methyl-5-methyluridine; 55486-09-4; 5-Methyl-2'-O-methyluridine; 2'-O-Methylribothymidine; 114952-97-5; 5-Me-2'-OMe Uridine; 1-((2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-methoxytetrahydrofuran-2-yl)-5-methylpyrimidine-2,4(1H,3H)-dione; 5,O2'-dimethyl-uridine
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.6731 mL | 18.3655 mL | 36.7309 mL | |
| 5 mM | 0.7346 mL | 3.6731 mL | 7.3462 mL | |
| 10 mM | 0.3673 mL | 1.8365 mL | 3.6731 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA