包装 | 价格(元) |
25ul (1 mM) | 电议 |
5x25ul (1 mM) | 电议 |
Physical Appearance | Solution |
Storage | Store at -20°C or below |
M.Wt | 1066.91 (free acid) |
Formula | C43H65N4O21P3(free acid) |
Synonyms | Digoxigenin-X-5-aminoallyl-2'-deoxyuridine-5'-triphosphate, Triethylammonium salt, DIG-11-dUTP |
Solubility | Soluble in sterile water |
Chemical Name | ((2R,3S,5R)-5-(5-((E)-3-(6-(2-(((3S,5R,8R,9S,10S,12R,13S,14S,17R)-12,14-dihydroxy-10,13-dimethyl-17-(5-oxo-2,5-dihydrofuran-3-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)acetamido)hexanamido)prop-1-en-1-yl)-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-y |
Canonical SMILES | O=C1OCC([C@H]2CC[C@@]3(O)[C@]2(C)[C@H](O)C[C@@]4([H])[C@@]3([H])CC[C@@]5([H])[C@]4(C)CC[C@H](OCC(NCCCCCC(NC/C=C/C6=CN([C@H]7C[C@H](O)[C@@H](COP(O)(OP(O)(OP(O)(O)=O)=O)=O)O7)C(NC6=O)=O)=O)=O)C5)=C1 |
运输条件 | 蓝冰运输或根据您的需求运输。 |
一般建议 | 为了使其更好的溶解,请用37℃加热试管并在超声波水浴中震动片刻。不同厂家不同批次产品溶解度各有差异,仅做参考。若实验所需浓度过大至产品溶解极限,请添加助溶剂助溶或自行调整浓度。溶液形式一般不宜长期储存,请尽快用完。 |
Digoxigenin-11-dUTP可以酶促掺入到DNA / cDNA中以替代其天然对应物dTTP。随后使用耦联辣根过氧化物酶(HRP),碱性磷酸酶(AP)或荧光染料的地高辛抗体检测得到的地高辛标记DNA / cDNA探针。通过连接在尿苷C5位上的11个原子的接头保证了最佳的底物特性和标记效率。
Digoxigenin-11-dUTP可以通过以下方法整合到DNA/cDNA中:
- 用Taq聚合酶进行PCR
- 用DNAse I / DNA聚合酶I进行Nick Translation
- 用Klenow exo-进行引物延伸
- 用末端脱氧核苷酸转移酶(TdT)标记寡核苷酸3'末端
- 用MMLV逆转录酶进行逆转录
Digoxigenin-11-dUTP可以通过以下方法整合到RNA中:
- 用末端脱氧核苷酸转移酶(TdT)标记3'末端
推荐的用于PCR和Nick Translation的Digoxigenin-11-dUTP / dTTP比例:35%地高辛-11-dUTP / 65%dTTP
请注意:Digoxigenin-11-dUTP的最佳终浓度取决于应用和实验条件。为获得最佳产品收率和高掺入率,建议单独优化Digoxigenin-11-dUTP / dTTP比率
参考文献:
[1] Anderson et al. (2005) Incorporation of reporter-labeled nucleotides by DNA polymerases. Biotechniques 38:257.
[2] Jackson et. al (1991) Detection of Shiga Toxin-Producing Shigella dysenteriae Type 1 and Escherichia coli by Using Polymerase Chain Reaction with Incorporation of Digoxigenin-11-dUTP. J Clin Microbiol. 29 (9):1910.
[3] Dauwerse et al. (1999) Two-colour FISH detection of the inv (16) in interphase nuclei of patients with acute myeloid leukemia. Br J Haematol 106:111.
[4] Wiegant et al. (2008) Probe Labeling and Fluorescence In Situ Hybridization. Current Protocols in Cytometry: Unit 8.3.
[5] Schmitz et al. (2008) Nonradioactive labeling of oligonucleotides in vitro with the hapten digoxigenin by tailing with terminal transferase. Anal Biochem 199:222.
[6] Grimmond et al. (2001) Expression Profiling with cDNA Microarray's: A User's Perspective and Guide. In: DNA Microarrays: Gene Expression Applications (Jordan). Springer Verlag Berlin Heidelberg.
[7] Rosemeyer et al. (1995) Nonradioactive 3'-end-labeling of RNA molecules of different lengths by terminal deoxynucleotidyltransferase Anal Biochem 224:446.