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In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
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.
References:[1] Schmitzet al. (1991) Nonradioactive labeling of oligonucleotides in vitro with the hapten digoxigenin by tailing with terminal transferase. Anal Biochem199:222.