Digoxigenin
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Preferred IUPAC name
4-[(1R,3aS,3bR,5aR,7S,9aS,9bS,11R,11aS)-3a,7,11-Trihydroxy-9a,11a-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-1-yl]furan-2(5H)-one | |
Identifiers | |
ChEBI | |
ChEMBL | |
ChemSpider | |
ECHA InfoCard | 100.015.279 |
PubChem CID
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UNII | |
CompTox Dashboard (EPA)
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Properties | |
C23H34O5 | |
Molar mass | 390.51 g/mol |
log P | 2.57510[1] |
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa). | |
what is ?) | (|
Infobox references | |
Digoxigenin (DIG) is a steroid found exclusively in the flowers and leaves of the plants Digitalis purpurea, and Digitalis lanata (foxgloves), where it is attached to sugars, to form the glycosides (e.g. Lanatoside C).[2]
Use in biotechnology[]
Digoxigenin is a hapten, a small molecule with high antigenicity, that is used in many molecular biology applications similarly to other popular haptens such as 2,4-Dinitrophenol, biotin, and fluorescein. Typically, digoxigenin is introduced chemically (conjugation) into biomolecules (proteins, nucleic acids) to be detected in further assays. Kd of the digoxigenin-antibody interaction has been estimated at ~12 nM [3] (compare to Kd~0.1pM for the biotin-streptavidin interaction[4]).
DIG-binding proteins. Tinberg et al. designed artificial proteins that bind DIG. Their best binder, DIG10.3, was a 141 amino acid protein that bound DIG with a dissociation constant (Kd) of 541 (+/- 193) pM.[5]
Anti-digoxigenin antibodies with high affinities and specificity are used in a variety of biological immuno-assays (e.g. ELISA). The antibodies are labeled with dyes, enzymes or fluorescence, directly or secondarily, for visualization and detection.
Digoxigenin is thus an all-purpose immuno-tag, and in particular a standard immunohistochemical marker for in situ hybridization.[6][7] In this case it is conjugated to a single species of RNA nucleoside triphosphate (typically uridine), which is then incorporated into RNA (a "riboprobe") as it is synthesized by the cellular machinery.
It allows to make :
- sensitive non-radioactive in situ hybridization probes to detect nucleic acids in plants, able to detect 1 µg of plasmid DNA.[8]
- peptide-DIG conjugates, i.e. bradykinin assay by very sensitive chemiluminescence immunoassays.[9]
- fluorescent and DIG-labeled tracers for competitive immunoassays, i.e. to limit detect digoxin, a drug used to cure cardiac arrhythmia, down to 0.2 ng mL−1.[10]
- Digoxigenin may be conjugated to sugars to study glycosylation events,[11] even in biological systems.
See also[]
References[]
- ^ "Digoxigenin". Material Data Safety Sheet. ChemSrc.
- ^ Polya G (2003). Biochemical Targets of Plant Bioactive Compounds. New York: CRC Press. ISBN 978-0415308298.
- ^ Tetin SY, Matayoshi ED (August 2002). "Measuring antibody affinity and performing immunoassay at the single molecule level". Analytical Biochemistry. 307 (1): 84–91. doi:10.1016/S0003-2697(02)00011-8.
- ^ Duan X. "Quantification of the affinities and kinetics of protein interactions using silicon nanowire biosensors". Nature Nanotechnology. doi:10.1038/nnano.2012.82. PMC 4180882.
- ^ Tinberg CE, Khare SD, Dou J, Doyle L, Nelson JW, Schena A, Jankowski W, Kalodimos CG, Johnsson K, Stoddard BL, Baker D (September 2013). "Computational design of ligand-binding proteins with high affinity and selectivity". Nature. 501 (7466): 212–216. doi:10.1038/nature12443. PMC 3898436. PMID 24005320.
- ^ Eisel D, Grünewald-Janho S, Krushen B, eds. (2002). DIG Application Manual for Nonradioactive in situ Hybridization (3rd ed.). Penzberg: Roche Diagnostics.
- ^ Hauptmann G, Gerster T (August 1994). "Two-color whole-mount in situ hybridization to vertebrate and Drosophila embryos". Trends in Genetics. 10 (8): 266. doi:10.1016/0168-9525(90)90008-T. PMID 7940754.
- ^ Hart SM, Basu C (April 2009). "Optimization of a digoxigenin-based immunoassay system for gene detection in Arabidopsis thaliana" (pdf). Journal of Biomolecular Techniques. 20 (2): 96–100. PMC 2685603. PMID 19503620.
- ^ Décarie A, Drapeau G, Closset J, Couture R, Adam A (1994). "Development of digoxigenin-labeled peptide: application to chemiluminoenzyme immunoassay of bradykinin in inflamed tissues". Peptides. 15 (3): 511–8. doi:10.1016/0196-9781(94)90214-3. PMID 7937327.
- ^ Mayilo S, Ehlers B, Wunderlich M, Klar TA, Josel HP, Heindl D, Nichtl A, Kürzinger K, Feldmann J (July 2009). "Competitive homogeneous digoxigenin immunoassay based on fluorescence quenching by gold nanoparticles". Analytica Chimica Acta. 646 (1–2): 119–22. doi:10.1016/j.aca.2009.05.023. PMID 19523564.
- ^ Goodarzi MT, Rafiq M, Turner G (May 1995). "An improved multiwell immunoassay using digoxigenin-labelled lectins to study the glycosylation of purified glycoproteins". Biochemical Society Transactions. 23 (2): 168S. doi:10.1042/bst023168s. PMID 7672194.
- Cardenolides
- Triols