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通过基于5-EdU的铜催化点击化学可视化斑马鱼的从头DNA合成。斑马鱼幼鱼在受精后4天（dpf）用10mM的5-乙炔基-2'-脱氧尿苷（EdU）处理。在5 dpf（追踪24小时）时，根据分步方案，使用CuAAC细胞反应缓冲液试剂盒（基于BTTAA）和AZDye647-吡啶甲基-叠氮化物对幼鱼进行固定染色。使用的转基因品系：Tg(fabp10a:H2B-mGL) 和 Tg(tp1:H2B-mCherry)肝细胞：绿色 响应性胆管细胞：红色 5-乙炔基-2'-脱氧尿苷：白色
For general laboratory use.
Shipping: shipped at ambient temperature
Storage Conditions: store at -20 °C
store dry and under inert gas
Short term exposure (up to 1 week cumulative) to ambient temperature possible.Shelf Life: 12 months after date of delivery
Molecular Formula: C11H12N2O5
Molecular Weight: 252.23 g/mol
Exact Mass: 252.07 g/mol
CAS#: 61135-33-9
Purity: ≥ 98 % (HPLC)
Form: solid
Color: white to off-white
Solubility: DMSO, up to 200 mM (at room temperature) in 1 x PBS or water by heating the obtained suspension for 1 minute by 70 °C
Spectroscopic Properties: λmax 288 nm, ε 12.0 L mmol-1 cm-1 (Tris-HCl pH 7.5)
Applications:
DNA synthesis monitoring[1-5]Description:
5-EdU (5-Ethynyl-2'-deoxyuridine) can be used as a replacement for BrdU (5-Bromo-2'-deoxyuridine) to measure de novo DNA synthesis during the S-phase of the cell cycle. 5-EdU is cell permeable and incorporates into replicating DNA instead of its natural analog thymidine. The resulting ethynyl-functionalized DNA can subsequently be detected via Cu(I)-catalyzed click chemistry that offers the choice to introduce a Biotin group (via Azides of Biotin) for subsequent purification tasks or a fluorescent group (via Azides of fluorescent dyes) for subsequent microscopic imaging [1-5].Presolski et al.[6] and Hong et al.[7] provide a general protocol for Cu(I)-catalyzed click chemistry reactions that may be used as a starting point for the set up and optimization of individual assays.
[1] Salic et al. (2008) A chemical method for fast and sensitive detection of DNA synthesis in vivo. Proc.Natl. Acad. Sci. USA 105: 2415.
[2] Hamelik et al. (2009) Click-iTTM Assay with Improved DNA Distribution Histograms. Cytometry A 75A:862.
[3] Diermeier-Daucher et al. (2009) Cell Type Specific Applicability of 5-Ethynyl-2’-deoxyuridine (EdU) for Dynamic Proliferation Assessment in Flow Cytometry. Cytometry A 75A:535
[4] Buck et al. (2008) Detection of S-Phase cell cycle progression using 5’-ethynyl-2’-deoxyuridine incorporation with click chemistry, an alternative to using 5’-bromo-2’-deoxyuridine. Biotechniques 44(7):927.
[5] Limsirichaikul et al. (2009) A rapid non-radioactive technique for measurement of repair synthesis in primary human fibroblasts by incorporation of ethynyl deoxyuridine (EdU). Nucleic Acids Res. 37(4):e31.
[6] Zeng et al. (2010) Evaluation of 5-ethynyl-2′-deoxyuridine staining as a sensitive and reliable method for studying cell proliferation in the adult nervous system. Brain Research. 1319:21.
[7] Warren et al. (2009) Chick embryo proliferation studies using EdU labeling. Dev Dyn. 238(4):944.[8] Monzack et al. (2012) A time course investigation of the statin paradox among valvular interstitial cell phenotypes. American Journal of Physiology. 303(7):H903.
[9] Gouge et al. (2010) Detection of S Phase in multiple Drosophila tissues utilizing the EdU labeling technique. Dros. Inf. Serv. 93: 213.
[10] Kotogány et al. (2010) A rapid and robust assay for detection of S-phase cell cycle progression in plant cells and tissues by using ethynyl deoxyuridine. Plant Methods 6: 5.
[11] Rieder et al. (2014) Alkene-Tetrazine Ligation for Imaging Cellular DNA. Angew. Chem. Int. Ed. 53: 9168. -
产品组分
内容
型号
规格 储存温度
5-乙炔基-2'-脱氧尿苷(5-EdU)
CLKN001-01 25mg
-20 °C
5-乙炔基-2'-脱氧尿苷(5-EdU)
CLKN001-02 100mg
-20 °C 5-乙炔基-2'-脱氧尿苷(5-EdU)
CLKN001-03 500mg
-20 °C 5-乙炔基-2'-脱氧尿苷(5-EdU)
CLKN001-04 5g
-20 °C 操作手册
1 1 常温
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注意事项
保存建议 厂家推荐蓝冰运输。当您收到产品后，按照说明书建议保存于-20°C。 -
FAQ
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Selected References:
[1] Salic et al. (2008) A chemical method for fast and sensitive detection of DNA synthesis in vivo. Proc. Natl. Acad. Sci. USA 105 (7):2415.
[2] Li et al. (2010) Fluorogenic click reaction for labeling and detection of DNA in proliferating cells. Biotechniques 49 (1):525.
[3] Chehrehasa et al. (2009) EdU, a new thymidine analogue for labelling proliferating cells in the nervous system. J. Neurosci. Methods 177:122.
[4] Limsirichaikul et al. (2009) A rapid non-radioactive technique for measurement of repair synthesis in primary human fibroblasts by incorporation of ethynyl deoxyuridine (EdU). Nucleic Acids Res. 37 (4):e31.
[5] Buck et al. (2008) Detection of S-phase cell cycle progression using 5-ethynyl-2'-deoxyuridine incorporation with click chemistry, an alternative to using 5-bromo-2'-deoxyuridine antibodies. Biotechniques 44 (7):927.
[6] Presolski et al. (2011) Copper-Catalyzed Azide-Alkyne Click Chemistry for Bioconjugation. Current Protocols in Chemical Biology 3:153.
[7] Hong et al. (2011) Analysis and Optimization of Copper-Catalyzed Azide-Alkyne Cycloaddition for Bioconjugation. Angew. Chem. Int. Ed. 48:9879.The nascent RNA labelling compound 5-ethynyl uridine (EU) integrates into DNA in some animals.
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