Digoxigenin-11-dUTP
Digoxigenin-11-dUTP is enzymatically incorporated into DNA/cDNA as substitute for its natural counterpart dTTP. The resulting Digoxigenin-labeled DNA/cDNA probes are subsequently detected using Digoxigenin-antibodies conjugated with horseradish peroxidase (HRP), alkaline phosphatase (AP) or a fluorescent dye. Optimal substrate properties and thus labeling efficiency is ensured by an 11-atom linker attached to the C5 position of uridine.
Digoxigenin-11-dUTP can be incorporated into DNA/cDNA by
- PCR with Taq polymerase
- Nick Translation with DNAse I/ DNA Polymerase I
- Primer Extension with Klenow exo-
- 3’-End Labeling with Terminal deoxynucleotidyl Transferase (TdT)
- Reverse Transcription with MMLV Reverse Transcriptase
Digoxigenin-11-dUTP can be incorporation into RNA by
- 3’-End Labeling with Terminal deoxynucleotidyl Transferase (TdT)
Recommended Digoxigenin-11-dUTP/dTTP ratio for PCR and Nick Translation: 35% Digoxigenin-11-dUTP/ 65% dTTP
Please note: The optimal final concentration of Digoxigenin-11-dUTP may depending on the application and assay conditions. For optimal product yields and high incorporation rates an individual optimization of the Digoxigenin-11-dUTP/dTTP ratio is recommended
References:
[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.
| 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 |
| SDF | Download SDF |
| 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 |
| Shipping Condition | Small Molecules with Blue Ice, Modified Nucleotides with Dry Ice. |
| General tips | We do not recommend long-term storage for the solution, please use it up soon. |
Quality Control & MSDS
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Purity = 98.00%
- COA (Certificate Of Analysis)
- MSDS (Material Safety Data Sheet)
Chemical structure
