Digoxigenin-11-ddUTP can be used as a substrate for: Terminal Transferase, DNA polymerase I (holoenzyme and Klenow fragment), T4 and T7 DNA polymerase or Taq DNA polymerase and reverse transcriptase (e.g., Transcriptor). It is preferentially used for 3'-end labeling of oligonucleotides with Terminal Transferase, recombinant. The DIG-labeled oligonucleotide can be used as a hybridization probe for: DNA and RNA transferring, Colony and plaque screening, in situ hybridization and so on. The resulting Digoxigenin-labeled oligonucleotide 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.
Recommended molar ratio of Digoxigenin-11-ddUTP to free 3'-OH groups: 10:1.
In addition to common hybridization techniques, DIG labeled oligonucleotides are specifically useful for screening expression libraries for sequence specific DNA-binding proteins such as transcription factors.
Please note: The optimal final concentration and molar ratio of Digoxigenin-11-ddUTP to free 3'-OH groups may depending on the application and assay conditions. For optimal incorporation rates an individual optimization of the Digoxigenin-11-ddUTP concentration and molar ratio is recommended.
 Schmitzet al. (1991) Nonradioactive labeling of oligonucleotides in vitro with the hapten digoxigenin by tailing with terminal transferase. Anal Biochem199:222.
|Storage||Desiccate at -20°C|
|M.Wt||1048.90 g/mol (free acid)|
|Formula||C43H63N4O20P3 (free acid)|
|Synonyms||Digoxigenin-3-O-methylcarbonyl-ε-aminocaproyl-[5-(3-aminopropargyl) -2',3'-dideoxyuridine-5'-triphosphate], Triethylammonium salt, Dig-11-ddUTP|
|Solubility||Soluble in sterile water|
|Canonical SMILES||O=C1OCC([[email protected]]2CC[[email protected]@]3(O)[[email protected]]2(C)[[email protected]](O)C[[email protected]@]4([H])[[email protected]@]3([H])CC[[email protected]@]5([H])[[email protected]]4(C)CC[[email protected]](OCC(NCCCCCC(NCC#CC6=CN([[email protected]]7CC[[email protected]@H](COP(O)(OP(O)(OP(O)(O)=O)=O)=O)O7)C(NC6=O)=O)=O)=O)C5)=C1|
|Shipping Condition||Evaluation sample solution : ship with blue ice.All other available size: ship with RT , or blue ice upon request|
|General tips||For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.|