Vinyl-containing nucleosides can be labeled with Tetrazine-tagged reporter molecules via Copper-free Tetrazine-Alkene Ligation that allows to introduce a Biotin group for subsequent purification (via Tetrazine-containing Biotinylation Reagents) or a fluorescent group for subsequent microscopic imaging (via Tetrazine-containing Fluorescent Dyes).

They are suitable for DNA synthesis monitoring or RNA synthesis monitoring, respectively.

Methylcyclopropene-containing monosaccharides can be used to attach methylcyclopropene groups to glycoconjugates via metabolic labeling ^[1-3] followed by subsequent labeling with Tetrazine-containing detection reagents via Cu(I)-free Tetrazine-Cyclopropene Ligation^[4].

A number of CLICKable monosaccharides are available for metabolic glycan labeling.

Selected References

[1] Spaete et al. (2014) Expanding the scope of cyclopropene reporters for the detection of metabolically engineered glycoproteins by Diels–Alder reactions. Beilstein J. Org. Chem. 10:2235.
[2] Spaete et al. (2014) Rapid labeling of metabolically engineered cell-surface glycoconjugates with a carbamate-linked cyclopropene reporter. Bioconj. Chem. 25:147.
[3] Patterson et al. (2014) Improved cyclopropene reporters for probing protein glycosylation. Mol. BioSyst. 10:1693.
[4] Patterson et al. (2012) Functionalized cyclopropenes as bioorthogonal chemical reporters. J. Am. Chem. Soc. 134:18643.

Bifunctional TCO-containing Reagents

Selected References:

Selvaraj et al. (2013) trans-Cyclooctene – a stable voracious dienophile for bioorthogonal labeling. Current Opinion in Chemical Biology 17:753.

Dibenzocyclooctyne (DBCO)^*-containing fluorescent dyes can be used for the fluorescent labeling of Azide-tagged biomolecules via Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC). Both Standard Dyes (e.g. Cy3, Tamra or Texas Red) and novel Alternative Dyes have been thoroughly selected to cover the whole UV-Vis spectrum.

^*DBCO is also known as ADIBO (= Azadibenzocyclooctyne)

Table 1: Available DBCO-containing Fluorescent Dyes

Dibenzocyclooctyne (DBCO)^*-containing (desthio) biotinylation reagents can be used for the labeling of Azide-tagged biomolecules via Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC).

A number of (desthio) biotinylation reagents with

• • different solubility characteristics (PEG- and Sulfo-linker for increased molecule solubility)

• different cleavability characteristics (chemically or photocleavable linke)

are available to fit specific application & molecule requirements.

^*DBCO is also known as ADIBO (= Azadibenzocyclooctyne) or DIBAC (= Dibenzoazacyclooctyne)

Table 1: Available DBCO-containing (Desthio) Biotinylation Reagents

Dibenzocyclooctyne (DBCO)^*-containing FLAG tag reagents can be used for the introduction of a FLAG-tag to Azide-tagged biomolecules via Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC). Molecule solubility and efficient FLAG-tag detection is ensured by the integrated PEG-linker.

^*DBCO is also known as ADIBO (= Azadibenzocyclooctyne) or DIBAC (= Dibenzoazacyclooctyne)

Selected References

[1] Hopp et al. (1988) A Short Polypeptide Marker Sequence Useful for Recombinant Protein Identification and Purification. Bio/Technology 6:1204.
[2] Einhauer et al. (2001) The FLAG peptide, a versatile fusion tag for the purification of recombinant proteins. J. Biochem. Biophys. Methods 49:455.
[3] Brizzard et al. (1997) Epitope tagging of recombinant proteins. Current Protocols in Neuroscience:5.8.1.

Dibenzocyclooctyne (DBCO)^*-containing bifunctional reagents can be used for the introduction of

• a functional group to Azide-labeled molecules via Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC)
• an DBCO group to various targets e.g. via NHS Ester- or Maleimide-based chemistry

^*DBCO is also known as ADIBO (= Azadibenzocyclooctyne) or DIBAC (= Dibenzoazacyclooctyne)

Table 1: Available DBCO-containing Bifunctional Reagents

Dibenzocyclooctyne (DBCO)^*-modified polyethylene glycol (PEG) polymers can be used for the introduction of a PEG moiety (“PEGylation”) to Azide-tagged biomolecules via Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC), respectively.

PEGylation of biomolecules such as proteins increases their water solubility and often improves their pharmacokinetic properties e.g. by a reduced immunogenicity and antigenicity.

^*DBCO is also known as ADIBO (= Azadibenzocyclooctyne) or DIBAC (= Dibenzoazacyclooctyne)

Dibenzocyclooctyne (DBCO)^*-containing nucleotides can be attached to Azide-tagged biomolecules via Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC).

Search for nucleotides that are suitable for enzymatic click functionalization of DNA or enzymatic click functionalization of RNA

^*DBCO is also known as ADIBO (= Azadibenzocyclooctyne) or DIBAC (= Dibenzoazacyclooctyne)

Dibenzocyclooctyne (DBCO)^*-containing nucleosides can be attached to Azide-tagged biomolecules via Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC).

^*DBCO is also known as ADIBO (= Azadibenzocyclooctyne) or DIBAC (= Dibenzoazacyclooctyne)

Dibenzocyclooctyne (DBCO)^* Agarose is suitable to covalently capture Azide-tagged biomolecules via Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC).

The proteins of interest need to be metabolically, enzymatically or chemically azide-tagged. Subsequently, the DBCO Agarose containing the covalently attached proteins can be washed with high stringency, virtually eliminating any non-specifically bound proteins.

^*DBCO is also known as ADIBO (= Azadibenzocyclooctyne) or DIBAC (= Dibenzoazacyclooctyne)

Dibenzocyclooctyne (DBCO)^* Magnetic Beads are suitable to covalently capture Azide-tagged biomolecules onto magnetic beads via Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC).

The proteins of interest need to be metabolically, enzymatically or chemically azide-tagged. Subsequently, the DBCO Magnetic Beads containing the covalently attached proteins can be washed with high stringency, virtually eliminating any non-specifically bound proteins.

^*DBCO is also known as ADIBO (= Azadibenzocyclooctyne) or DIBAC (= Dibenzoazacyclooctyne)

Alkyne-containing Fluorescent dyes can be used for the fluorescent labeling of Azide-tagged molecules via a Cu(I)-catalyzed Azide-Alkyne Click Chemistry reaction (CuAAC).

Standard Dyes (e.g. Cy3, Tamra or Texas Red) have been thoroughly selected to cover the whole UV-Vis spectrum.

Table 1: Available Alkyne-containing Fluorescent Dyes

Alkyne-containing (desthio) biotinylation reagents can be used for the labeling of Azide-tagged biomolecules via a Cu(I)-catalyzed Azide-Alkyne Click Chemistry reaction (CuAAC).

A number of (desthio) biotinylation reagents with

• • improved solubility & detection characteristics (PEG-linker for increased molecule solubility & optimal streptavidin binding)

• different cleavability characteristics (chemically or photocleavable linker)

are available to fit specific application & molecule requirements.

Table 1: Available Alkyne-containing (Desthio) Biotinylation Reagents

Selected References

[1] Hopp et al. (1988) A Short Polypeptide Marker Sequence Useful for Recombinant Protein Identification and Purification. Bio/Technology 6:1204.
[2] Einhauer et al. (2001) The FLAG peptide, a versatile fusion tag for the purification of recombinant proteins. J. Biochem. Biophys. Methods 49:455.
[3] Brizzard et al. (1997) Epitope tagging of recombinant proteins. Current Protocols in Neuroscience:5.8.1.

Alkyne-containing reagents can be used for the introduction of

• a functional group to Azide-labeled molecules via a Cu(I)-catalyzed Azide-Alkyne Click Chemistry reaction (CuAAC)
• an Alkyne group to various targets e.g. via NHS Ester- or Maleimide-based chemistry

Table 1: Available Alkyne-containing Bifunctional Reagents

Alkyne-containing nucleotides can be attached to Azide-tagged biomolecules or incorporated into DNA, RNA or proteins followed by subsequent labeling with Azide-containing reporter molecules via a Cu(I)-catalyzed Azide-Alkyne Click Chemistry reaction (CuAAC).

Search for nucleotides that are suitable for enzymatic click functionalization of DNA or enzymatic click functionalization of RNA

Alkyne-containing nucleosides can be labeled with Azide-containing reporter molecules via a Cu(I)-catalyzed Azide-Alkyne Click Chemistry reaction (CuAAC).

A number of cell-permeable CLICK-functionalized nucleosides are suitable for:

DNA synthesis monitoring,
RNA synthesis monitoring,
the analysis of poly A tail dynamics or
Protein synthesis monitoring.

Alkyne-containing amino acids can be labeled with Azide-containing reporter molecules via a Cu(I)-catalyzed Azide-Alkyne Click Chemistry reaction (CuAAC).

A number of cell-permeable Click-functionalized amino acids are randomly incorporated instead of methionine during translation and are therefore suitable for residue selective protein synthesis monitoring.

Alkyne Magnetic Beads are suitable to covalently capture Azide-tagged biomolecules onto magnetic beads by a Cu(I)-catalyzed Azide-Alkyne Click chemistry reaction (CuAAC).

The proteins of interest need to be metabolically, enzymatically or chemically azide-tagged. Subsequently, the Alkyne Magnetic Beads containing the covalently attached proteins can be washed with high stringency, virtually eliminating any non-specifically bound proteins.

Tetrazine-containing fluorescent dyes are suitable for fluorescent labeling of Cyclooctene-labeled molecules via Cu-free Click Chemistry. Both Standard Dyes (e.g. Cy3 or Tamra) and novel Alternative Dyes have been thoroughly selected to cover the whole UV-Vis spectrum.

Two Tetrazine versions with different reactivities and stability characteristics are available to meet specific application requirements. Tetrazine reagents are the ideal choice if a rapid reaction kinetic is the key aspect whereas 6-Methyl-Tetrazine reagents are ideally suited if an improved chemical stability is required¹.

Table 1: Available Tetrazine-containing Fluorescent Dyes

Selected References

[1] Karver et al. (2011) Synthesis and Evaluation of a Series of 1,2,4,5-Tetrazines for Bioorthogonal Conjugations. Am. Chem. Soc. 22:2263.

Tetrazine-containing biotinylation reagents can be used for the labeling of cyclooctene (e.g. TCO) via Cu(I)-free Click Chemistry.

Two Tetrazine versions with different reactivities and stability characteristics are available to meet specific application requirements. Tetrazine reagents are the ideal choice if a rapid reaction kinetic is the key aspect whereas 6-Methyl-Tetrazine reagents are ideally suited if an improved chemical stability is required ^[1]. Both biotinylation reagents contain a PEG linker for increased molecule solubility and optimal streptavidin binding.

Selected References

[1] Karver et al. (2011) Synthesis and Evaluation of a Series of 1,2,4,5-Tetrazines for Bioorthogonal Conjugations. Am. Chem. Soc. 22:2263.

Tetrazine-containing reagents are suitable for the introduction of

• a functional group (e.g. Amine or NHS Ester) to Cyclooctene-labeled molecules via Cu-free Click Chemistry
• an Tetrazine group to various targets e.g. via NHS Ester- or Maleimide-based chemistry

Two Tetrazine versions with different reactivities and stability characteristics are available to meet specific application requirements. Tetrazine reagents are the ideal choice if a rapid reaction kinetic is the key aspect whereas 6-Methyl-Tetrazine reagents are ideally suited if an improved chemical stability is required^[1].

Table 1: Available Tetrazine-containing Bifunctional Reagents

Selected References

[1] Karver et al. (2011) Synthesis and Evaluation of a Series of 1,2,4,5-Tetrazines for Bioorthogonal Conjugations. Am. Chem. Soc. 22:2263.

Single Substances

Reaction Kits

Selected references:

[1] Presolski et al. (2011) Copper-Catalyzed Azide-Alkyne Click Chemistry for Bioconjugation. Current Protocols in Chemical Biology 3:153.
[2] Hong et al. (2011) Analysis and Optimization of Copper-Catalyzed Azide-Alkyne Cycloaddition for Bioconjugation. Angew. Chem. Int. Ed. 48:9879.
[3] Besanceney-Webler et al. (2011) Increasing the Efficiacy of Bioorthogonal Click Reactions for Bioconjugation: A Comparative Study. Angew. Chem. Int. Ed. 50:8051.
[4] Uttamapinant et al. (2012) Fast, Cell-Compatible Click Chemistry with Copper-Chelating Azides for Biomolecular Labeling. Angew. Chem. Int. Ed. 51:5852.

(Picolyl-) Azides of fluorescent dyes can be used for the fluorescent labeling of terminal Alkyne and strained Alknye (e.g DBCO)-labeled molecules via Cu(I)-catalyzed Alknye-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction, respectively.

Standard Dyes such as Cy3, Tamra or Texas Red have been thoroughly selected to cover the whole UV-Vis spectrum. Novel Alternative Dyes are available as well.

Table 1: Available (Picolyl-) Azide-containing Fluorescent Dyes

(Picolyl-) Azides of (desthio) biotinylation reagents can be used for the labeling of terminal Alkyne- and strained Alkyne (e.g. DBCO)-labeled biomolecules via Cu(I)-catalyzed Alknye-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction, respectively.

A number of (desthio) biotinylation reagents with

• • different solubility characteristics (PEG-linker for increased molecule solubility)

• different cleavability characteristics (chemically or photocleavable linker)

are available to fit specific application & molecule requirements.

Table 1: Available (Picolyl) Azide-containing (Desthio) Biotinylation Reagents

Azide-containing FLAG tag reagents can be used for the introduction of a FLAG-tag to terminal Alkyne and strained Alkyne (e.g. DBCO)-labeled biomolecules via Cu(I)-catalyzed Alknye-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction, respectively.

Molecule solubility and efficient FLAG-tag detection is ensured by the integrated PEG-linker.

Selected References

[1] Hopp et al. (1988) A Short Polypeptide Marker Sequence Useful for Recombinant Protein Identification and Purification. Bio/Technology 6:1204.
[2] Einhauer et al. (2001) The FLAG peptide, a versatile fusion tag for the purification of recombinant proteins. J. Biochem. Biophys. Methods 49:455.
[3] Brizzard et al. (1997) Epitope tagging of recombinant proteins. Current Protocols in Neuroscience:5.8.1.

Azide-containing bifunctional reagents are suitable for the introduction of

• a functional group to terminal Alkyne-tagged or strained Alkyne (e.g. DBCO)-tagged molecules via Cu(I)-catalyzed Alknye-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction, respectively
• an Azide group to various targets e.g. via NHS Ester- or Maleimide-based chemistry

Table 1: Available Azide-containing Bifunctional Reagents

Azide-containing nucleosides can be labeled with terminal Alkyne or strained Alkyne (e.g. DBCO)-tagged reporter molecules via a Cu(I)-catalyzed Alknye-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction, respectively.

A number of cell-permeable CLICK-functionalized nucleosides are suitable for:

DNA synthesis monitoring,
RNA synthesis monitoring,
the analysis of poly A tail dynamics or
Protein synthesis monitoring.

Azides of amino acids can be labeled with terminal Alkyne or strained Alkyne (e.g. DBCO)-tagged reporter molecules via a Cu(I)-catalyzed Alkyne-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction, respectively.

A number of cell-permeable Click-functionalized amino acids are randomly incorporated instead of methionine during translation and are therefore suitable for residue selective protein synthesis monitoring.

Azides of monosaccharides can be labeled with terminal Alkyne or strained Alkyne (e.g. DBCO)-tagged reporter molecules via a Cu(I)-catalyzed Alkyne-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction, respectively.

Azide Agarose is suitable to covalently capture terminal Alkyne and strained Alkyne (e.g DBCO)-tagged biomolecules via a Cu(I)-catalyzed Alkyne-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction, respectively.

The proteins of interest need to be metabolically, enzymatically or chemically terminal Alkyne- or DBCO-tagged. Subsequently, the Azide Agarose containing the covalently attached proteins can be washed with high stringency, virtually eliminating any non-specifically bound proteins.

Azide Magnetic Beads are suitable to covalently capture terminal Alkyne and strained Alkyne (e.g. DBCO)-tagged biomolecules onto magnetic beads by via a Cu(I)-catalyzed Alkyne-Azide (CUAAC) or Cu(I)-free strain-promoted Alkyne-Azide Click Chemistry (SPAAC) reaction, respectively.

The proteins of interest need to be metabolically, enzymatically or chemically terminal Alkyne- or DBCO-tagged. Subsequently, the Azide Magnetic Beads containing the covalently attached proteins can be washed with high stringency, virtually eliminating any non-specifically bound proteins.