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{{cs1 config|name-list-style=vanc}}
{{chembox
{{chembox
| Watchedfields = changed
| verifiedrevid = 460786276
| verifiedrevid = 460786276
| Name =
|ImageFile=Digoxigenin acsv.svg
| ImageFile = Digoxigenin acsv.svg
|ImageSize=200px
| ImageSize = 200px
|IUPACName=3-[(3''S'',5''R'',8''R'',9''S'',10''S'',12''R'',13''S'',14''S'',17''R'')-3,12,14-trihydroxy-10,13-dimethyl-1,2,3,4,5,6,7,8,9,11,12,15,16,17-tetradecahydrocyclopenta[''a'']phenanthren-17-yl]-2''H''-furan-5-one
| IUPACName = 3β,12β,14-Trihydroxy-5β-card-20(22)-enolide
⚫
|OtherNames=
| SystematicName = 4-[(1''R'',3a''S'',3b''R'',5a''R'',7''S'',9a''S'',9b''S'',11''R'',11a''S'')-3a,7,11-Trihydroxy-9a,11a-dimethylhexadecahydro-1''H''-cyclopenta[''a'']phenanthren-1-yl]furan-2(5''H'')-one
⚫
|Section1={{Chembox Identifiers
⚫
| OtherNames =
⚫
| ChemSpiderID_Ref = {{chemspidercite|correct|chemspider}}
⚫
| Section1 = {{Chembox Identifiers
⚫
| ChemSpiderID_Ref = {{chemspidercite|correct|chemspider}}
| ChemSpiderID = 14728
| ChemSpiderID = 14728
| InChI = 1/C23H34O5/c1-21-7-5-15(24)10-14(21)3-4-17-18(21)11-19(25)22(2)16(6-8-23(17,22)27)13-9-20(26)28-12-13/h9,14-19,24-25,27H,3-8,10-12H2,1-2H3/t14-,15+,16-,17-,18+,19-,21+,22+,23+/m1/s1
| InChI = 1/C23H34O5/c1-21-7-5-15(24)10-14(21)3-4-17-18(21)11-19(25)22(2)16(6-8-23(17,22)27)13-9-20(26)28-12-13/h9,14-19,24-25,27H,3-8,10-12H2,1-2H3/t14-,15+,16-,17-,18+,19-,21+,22+,23+/m1/s1
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| CASNo_Ref = {{cascite|correct|CAS}}
| CASNo_Ref = {{cascite|correct|CAS}}
| CASNo=1672-46-4
| CASNo=1672-46-4
| ChEMBL_Ref = {{ebicite|correct|EBI}}
| UNII_Ref = {{fdacite|correct|FDA}}
| UNII = NQ1SX9LNAU
| ChEMBL_Ref = {{ebicite|correct|EBI}}
| ChEMBL = 1153
| ChEMBL = 1153
| PubChem=15478
| PubChem=15478
| ChEBI_Ref = {{ebicite|correct|EBI}}
| ChEBI_Ref = {{ebicite|correct|EBI}}
| ChEBI = 42098
| ChEBI = 42098

| SMILES = O=C/1OCC(=C\1)\[C@@H]2[C@@]4([C@](O)(CC2)[C@@H]5CC[C@@H]3C[C@@H](O)CC[C@]3(C)[C@H]5C[C@H]4O)C
}}
}}
|Section2={{Chembox Properties
| Section2 = {{Chembox Properties
| Formula=C<sub>23</sub>H<sub>34</sub>O<sub>5</sub>
| Formula=C<sub>23</sub>H<sub>34</sub>O<sub>5</sub>
| MolarMass=390.51 g/mol
| MolarMass=390.51 g/mol
| Appearance=
| Appearance=
| Density=
| Density=
| MeltingPt=
| MeltingPt=
| BoilingPt=
| BoilingPt=
| Solubility=
| Solubility=
| LogP = 2.57510<ref name="chemsrc">{{Cite web|url=https://www.chemsrc.com/en/cas/1672-46-4_409065.html|title=Digoxigenin | work = Material Data Safety Sheet | publisher = ChemSrc }}</ref>
}}
}}
|Section3={{Chembox Hazards
| Section3 = {{Chembox Hazards
| MainHazards=
| MainHazards=
| FlashPt=
| FlashPt=
| AutoignitionPt =
| Autoignition=
}}
}}
| Section4 =
| Section5 =
| Section6 =
}}
}}


'''Digoxigenin''' (DIG) is a [[steroid]] found exclusively in the flowers and leaves of the plants ''[[Digitalis purpurea]]'', ''[[Digitalis orientalis]]'' and ''[[Digitalis lanata]]'' (foxgloves), where it is attached to sugars, to form the [[Glycoside|glycosides]] (e.g. [[Lanatoside C]]).<ref>{{ cite book | author = Polya, G. | title = Biochemical Targets of Plant Bioactive Compounds | location = New York | publisher = CRC Press | year = 2003 | isbn = 978-0415308298 }}</ref>
'''Digoxigenin''' ('''DIG''') is a [[steroid]] found exclusively in the flowers and leaves of the plants ''[[Digitalis purpurea]]'', ''[[Digitalis orientalis]]'' and ''[[Digitalis lanata]]'' (foxgloves), where it is attached to sugars, to form the [[glycoside]]s (e.g. [[digoxin]], [[lanatoside C]]).<ref>{{ cite book | vauthors = Polya G | title = Biochemical Targets of Plant Bioactive Compounds | location = New York | publisher = CRC Press | year = 2003 | isbn = 978-0-415-30829-8 }}</ref>


== Use in biotechnology ==
== Uses in biotechnology ==


Digoxigenin is a [[hapten]], a small molecule with high [[immunogenic|immunogenicity]], that is used in many molecular biology applications similarly to other popular haptens such as DNP ([[2,4-Dinitrophenol|dinitrophenol]]), [[biotin]], and [[fluorescein]]. Typically, Digoxigenin is introduced chemically ([[bioconjugation|conjugation]]) into biomolecules (proteins, nucleic acids) to be detected in further assays. Anti-Digoxigenin [[antibody|antibodies]] with high [[Affinity (pharmacology)|affinities]] and specificity, are used in a variety of biological immuno-assays ([[ELISA]], ImmunoChemistry,...). Abs are labeled with dyes, enzymes or fluorescence, directly or secondarily, for visualization and detection.
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 ([[bioconjugation|conjugation]]) into biomolecules (proteins, nucleic acids) to be detected in further assays. K<sub>d</sub> of the digoxigenin-antibody interaction has been estimated at ~12 nM <ref>{{cite journal | vauthors = Tetin SY, Matayoshi ED | title = Measuring antibody affinity and performing immunoassay at the single molecule level | journal = Analytical Biochemistry| volume = 307| issue = 1| pages = 84–91| date = August 2002 | doi = 10.1016/S0003-2697(02)00011-8| pmid = 12137783 }}</ref> (compare to K<sub>d</sub>~0.1pM for the biotin-streptavidin interaction<ref>{{cite journal | vauthors = Duan X | title = Quantification of the affinities and kinetics of protein interactions using silicon nanowire biosensors | journal = Nature Nanotechnology| date = 2012 | volume = 7 | issue = 6 | pages = 401–407 | doi = 10.1038/nnano.2012.82| pmid = 22635097 | pmc = 4180882| bibcode = 2012NatNa...7..401D }}</ref>).


;DIG-binding proteins
⚫
Digoxigenin is thus an all-purpose immuno-tag, and in particular a standard [[immunohistochemistry|immunohistochemical]] marker for ''in situ'' hybridization.<ref>{{ cite book | editor = Eisel, D.; Grünewald-Janho, S.; Krushen, B. | title = DIG Application Manual for Nonradioactive ''in situ'' Hybridization | edition = 3rd | location = Penzberg | publisher = Roche Diagnostics | year = 2002 }}</ref><ref>{{ cite journal | author = Hauptmann, G.; Gerster, T. | title = Two-color whole-mount ''in situ'' hybridization to vertebrate and ''Drosophila'' embryos | journal = Trends in Genetics | year = 1994 | volume = 10 | issue = 8 | pages = 266 | doi = 10.1016/0168-9525(90)90008-T | pmid = 7940754 }}</ref> In this case it is conjugated to a single species of RNA [[nucleotide]] triphosphate (typically [[uridine]]), which is then incorporated into RNA (a "[[riboprobe]]") as it is synthesized by the cellular machinery.
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]] (K<sub>d</sub>) of 541 (+/- 193) pM.<ref>{{cite journal | vauthors = Tinberg CE, Khare SD, Dou J, Doyle L, Nelson JW, Schena A, Jankowski W, Kalodimos CG, Johnsson K, Stoddard BL, Baker D | title = Computational design of ligand-binding proteins with high affinity and selectivity | journal = Nature | volume = 501 | issue = 7466 | pages = 212–216 | date = September 2013 | pmid = 24005320 | pmc = 3898436 | doi = 10.1038/nature12443 | bibcode = 2013Natur.501..212T }}</ref>


Anti-digoxigenin [[antibody|antibodies]] with high [[Affinity (pharmacology)|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.
⚫
It allows to make :
⚫
* sensitive non-radioactive in situ hybridization probes to detect nucleic acids in plants, able to detect 1µg of plasmid DNA.<ref>{{ cite journal | author = Hart, S. M.; Basu, C. | title = Optimization of a Digoxigenin-Based Immunoassay System for Gene Detection in ''Arabidopsis thaliana'' | journal = Journal of Biomolecular Techniques | year = 2009 | volume = 20 | issue = 2 | pages = 96–100 | pmid = 19503620 | pmc = 2685603 | url = http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2685603/pdf/jbt-20-96.pdf | format = pdf }}</ref>
⚫
* peptides-DIG conjugates, i.e. Bradikinin assay by very sensitive chemiluminescence immunoassays.<ref>{{ cite journal | author = Décarie, A.; Drapeau, G.; Closset, J.; Couture, R.; Adam, A. | title = Development of a Digoxigenin-labeled Peptide: Application to a Chemiluminoenzyme Immunoassay of Bradykinin in Inflamed Tissues | journal = Peptides | year = 1994 | volume = 15 | issue = 3 | pages = 511–518 | doi = 10.1016/0196-9781(94)90214-3 | pmid = 7937327 }}</ref>
⚫
* 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<sup>-1</sup>.<ref>{{ cite journal | author = Mayilo, S.; Ehlers, B.; Wunderlich, M.; Klar, T. A.; Josel, H. P.; Heindl, D.; Nichtl, A.; Kürzinger, K.; Feldmann, J. | title = Competitive Homogeneous Digoxigenin Immunoassay Based on Fluorescence Quenching by Gold Nanoparticles | journal = Analytica Chimica Acta | year = 2009 | volume = 646 | issue = 1–2 | pages = 119–122 | doi = 10.1016/j.aca.2009.05.023 | pmid = 19523564 }}</ref>
⚫
* Digoxigenin may be conjugated to [[monosaccharide|sugars]] to study [[glycosylation]] events<ref>{{ cite journal | author = Goodarzi, M. T.; Rafiq, M.; Turner, G. | title = An Improved Multiwell Immunoassay Using Digoxigenin-Labelled Lectins to Study the Glycosylation of Purified Glycoproteins | journal = Biochemical Society Transactions | year = 1995 | volume = 23 | issue = 2 | pages = 168S | pmid = 7672194 }}</ref>, even in biological systems.


⚫
Digoxigenin is thus an all-purpose immuno-tag, and in particular a standard [[immunohistochemistry|immunohistochemical]] marker for [[in situ hybridization]].<ref>{{ cite book | veditors = Eisel D, Grünewald-Janho S, Krushen B | title = DIG Application Manual for Nonradioactive ''in situ'' Hybridization | edition = 3rd | location = Penzberg | publisher = Roche Diagnostics | year = 2002 }}</ref><ref>{{cite journal | vauthors = Hauptmann G, Gerster T | title = Two-color whole-mount in situ hybridization to vertebrate and Drosophila embryos | journal = Trends in Genetics | volume = 10 | issue = 8 | pages = 266 | date = August 1994 | pmid = 7940754 | doi = 10.1016/0168-9525(90)90008-T }}</ref> 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.
⚫
== References ==

{{reflist}}
⚫
It allows to make :
⚫
* sensitive non-radioactive in situ hybridization probes to detect nucleic acids in plants, able to detect 1&nbsp;μg of plasmid DNA.<ref>{{cite journal | vauthors = Hart SM, Basu C | title = Optimization of a digoxigenin-based immunoassay system for gene detection in Arabidopsis thaliana | journal = Journal of Biomolecular Techniques | volume = 20 | issue = 2 | pages = 96–100 | date = April 2009 | pmid = 19503620 | pmc = 2685603 }}</ref>
⚫
* peptide-DIG conjugates, i.e. bradykinin assay by very sensitive chemiluminescence immunoassays.<ref>{{cite journal | vauthors = Décarie A, Drapeau G, Closset J, Couture R, Adam A | title = Development of digoxigenin-labeled peptide: application to chemiluminoenzyme immunoassay of bradykinin in inflamed tissues | journal = Peptides | volume = 15 | issue = 3 | pages = 511–8 | year = 1994 | pmid = 7937327 | doi = 10.1016/0196-9781(94)90214-3 | s2cid = 19210640 }}</ref>
⚫
* fluorescent and DIG-labeled tracers for competitive immunoassays, i.e. to [[detection limit|limit detect]] digoxin, a drug used to cure cardiac arrhythmia, down to 0.2&nbsp;ng mL<sup>−1</sup>.<ref>{{cite journal | vauthors = Mayilo S, Ehlers B, Wunderlich M, Klar TA, Josel HP, Heindl D, Nichtl A, Kürzinger K, Feldmann J | title = Competitive homogeneous digoxigenin immunoassay based on fluorescence quenching by gold nanoparticles | journal = Analytica Chimica Acta | volume = 646 | issue = 1–2 | pages = 119–22 | date = July 2009 | pmid = 19523564 | doi = 10.1016/j.aca.2009.05.023 | bibcode = 2009AcAC..646..119M }}</ref>
⚫
* Digoxigenin may be conjugated to [[monosaccharide|sugars]] to study [[glycosylation]] events,<ref>{{cite journal | vauthors = Goodarzi MT, Rafiq M, Turner G | title = An improved multiwell immunoassay using digoxigenin-labelled lectins to study the glycosylation of purified glycoproteins | journal = Biochemical Society Transactions | volume = 23 | issue = 2 | pages = 168S | date = May 1995 | pmid = 7672194 | doi = 10.1042/bst023168s }}</ref> even in biological systems.


== See also ==
== See also ==
* [[Immunostaining]]
* [[Immunostaining]]


⚫
== References ==
⚫
[[Category:Cardenolides]]
{{Reflist}}


⚫
[[Category:Cardenolides]]
[[de:Digoxigenin]]
[[Category:Triols]]
[[fa:دیگوکسیژنین]]
[[fr:Digoxygénine]]
[[it:Digossigenina]]
[[ja:ジゴキシゲニン]]
[[pt:Digoxigenina]]

Latest revision as of 21:16, 10 May 2024

Digoxigenin
Names
IUPAC name
3β,12β,14-Trihydroxy-5β-card-20(22)-enolide
Systematic 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 Edit this at Wikidata
UNII
  • InChI=1S/C23H34O5/c1-21-7-5-15(24)10-14(21)3-4-17-18(21)11-19(25)22(2)16(6-8-23(17,22)27)13-9-20(26)28-12-13/h9,14-19,24-25,27H,3-8,10-12H2,1-2H3/t14-,15+,16-,17-,18+,19-,21+,22+,23+/m1/s1 checkY
    Key: SHIBSTMRCDJXLN-KCZCNTNESA-N checkY
  • InChI=1/C23H34O5/c1-21-7-5-15(24)10-14(21)3-4-17-18(21)11-19(25)22(2)16(6-8-23(17,22)27)13-9-20(26)28-12-13/h9,14-19,24-25,27H,3-8,10-12H2,1-2H3/t14-,15+,16-,17-,18+,19-,21+,22+,23+/m1/s1
    Key: SHIBSTMRCDJXLN-KCZCNTNEBN
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).
checkY verify (what is checkY☒N ?)

Digoxigenin (DIG) is a steroid found exclusively in the flowers and leaves of the plants Digitalis purpurea, Digitalis orientalis and Digitalis lanata (foxgloves), where it is attached to sugars, to form the glycosides (e.g. digoxin, lanatoside C).[2]

Uses in biotechnology

[edit]

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

[edit]

References

[edit]
  1. ^ "Digoxigenin". Material Data Safety Sheet. ChemSrc.
  2. ^ Polya G (2003). Biochemical Targets of Plant Bioactive Compounds. New York: CRC Press. ISBN 978-0-415-30829-8.
  3. ^ 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. PMID 12137783.
  4. ^ Duan X (2012). "Quantification of the affinities and kinetics of protein interactions using silicon nanowire biosensors". Nature Nanotechnology. 7 (6): 401–407. Bibcode:2012NatNa...7..401D. doi:10.1038/nnano.2012.82. PMC 4180882. PMID 22635097.
  5. ^ 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. Bibcode:2013Natur.501..212T. doi:10.1038/nature12443. PMC 3898436. PMID 24005320.
  6. ^ Eisel D, Grünewald-Janho S, Krushen B, eds. (2002). DIG Application Manual for Nonradioactive in situ Hybridization (3rd ed.). Penzberg: Roche Diagnostics.
  7. ^ 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.
  8. ^ Hart SM, Basu C (April 2009). "Optimization of a digoxigenin-based immunoassay system for gene detection in Arabidopsis thaliana". Journal of Biomolecular Techniques. 20 (2): 96–100. PMC 2685603. PMID 19503620.
  9. ^ 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. S2CID 19210640.
  10. ^ 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. Bibcode:2009AcAC..646..119M. doi:10.1016/j.aca.2009.05.023. PMID 19523564.
  11. ^ 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.
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