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Bromohexyl (5') Azide & Halotag Conjugation

picture of Bromohexyl (5') Azide & Halotag Conjugation

Modification : Bromohexyl/Azide (5')

Catalog Reference Number
Modification Code
5 Prime
3 Prime
Molecular Weight (mw)
Extinction Coeficient (ec)
Technical Info (pdf)
Absorbance MAX
Emission MAX
Absorbance EC

Click Chemistry

Catalog NoScalePrice
26-6923-0550 nmol$180.00
26-6923-02200 nmol$180.00
26-6923-011 umol$250.00
26-6923-032 umol$310.00
26-6923-1010 umol$936.00
26-6923-1515 umol$1,270.00

Bromohexyl modification is available as an Azide for Click Chemistry or as bromohexyl for Halotag protein conjugation. For Halotag protein conjugation a Spacer 18 modification should be added internally next to the 5'-Bromohexyl. Additional charge applies for Spacer 18 modification. Please specify in comments at time of order placement. Bromohexyl modification version has a setup charge of $250.00 for mild synthesis reagents per order. The azide version does not have additional charges

Click Chemistry Conjugation

The 5'-Bromohexyl modifier is used to make 5'-bromohexyl oligonucleotides, which are convertible into azidohexyl oligos on the synthesis column by treatment of the CPG with a solution of sodium azide and sodium iodide in dry DMF at 65 degree C. The CPG with the 5'-azido oligo is treated with ammonia affording 5'-azidohexyl-oligos in solution. 5'-azidohexyl oligos can be reacted with alkyne-end modified oligos using Click Chemistry (1,2) to form oligos with unique structures (branched, ringed, etc.).

Halotag Protein Oligo Conjugation

Click here for a validated Glen Research Protocol for Oligo Conjugation to Halo Tagged Protein

The strategy of small-molecule fluorescent labeling of genetically encoded proteins has become a popular alternative to GFP labeling.
Among the most widely used approaches is the HaloTag method developed by Promega, which utilizes a bacterial haloalkane dehalogenase. The enzyme removes halides from aliphatic hydrocarbons by a nucleophilic displacement mechanism to form a covalent ester linkage between the haloalkane and Asp106 in the enzyme. In the wild type haloalkane dehalogenase, the ester is quickly hydrolyzed by histidine 272 in the catalytic active site. However, by mutating the histidine to phenylalanine, the HaloTag variant renders the covalent ester bond stable toward hydrolysis.
Oligonucleotides should be synthesized with Bromohexyl at the 5' end with an adjacent internal Spacer 18 followed by the sequence of choice to be conjugated. Please note that for our online ordering system the addition of Spacer 18 modification is not automatic and should be added as an internal modification. For fluorescent detection the oligo can be labelled at the 3' end with a fluorophore.


Click Chemistry
1. Huisgen, R. Angew. Chem. Int. Ed. (1963), 2: 565-568.
2. Rostovtsev, V.V., Green, L.G., Fokin, V.V., Sharpless, K.B. A Stepwise Huisgen Cycloaddition Process: Copper(I)-Catalyzed Regioselective Ligation of Azides and Terminal Alkynes. Angew. Chem. Int. Ed. (2002), 41: 2596-2599.

Halotag Protein Conjugation
3. 1. Los, G. V.; Encell, L. P.; McDougall, M. G.; Hartzell, D. D.; Karassina, N.; Zimprich, C.; Wood, M. G.; Learish, R.; Ohana, R. F.; Urh, M.; Simpson, D.; Mendez, J.; Zimmerman, K.; Otto, P.; Vidugiris, G.; Zhu, J.; Darzins, A.; Klaubert, D. H.; Bulleit, R. F.; Wood, K. V. HaloTag: a novel protein labeling technology for cell imaging and protein analysis. ACS Chem. Biol., 2008, 3, 373-382
4. Vijay Singh, Shenliang Wang, and Eric T. Kool, Genetically Encoded Multispectral Labeling of Proteins with Polyfluorophores on a DNA Backbone. J. Am. Chem. Soc., 2013, 16, 6184-6191.
- Bromohexyl (5') Azide & Halotag Conjugation

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