Intended for the CuAAC reaction with proteins, protein labeling buffer protects biomolecules from being damaged by reactive
oxygen species. This reaction can be used for proteins with alkyne or azide groups, as well as cells
or components of cell lysates metabolically labeled with azide groups.
Azide is conjugated
to terminal alkyne, resulting in a five-membered heterocycle (1,2,3-triazole). Both groups (azide and
alkyne) are extremely rarely found in natural biomolecules, so the reaction is highly specific and
effective to handle various tasks.
The reaction proceeds in the
presence of copper (I) compounds and almost does not depend on pH. Optimized for operations with proteins,
protein labeling buffer contains a salt of copper (II) (a stable precursor of catalytically-active
copper (I) compounds), triethylammonium acetate pH 6.8, water-soluble THPTA ligand, and aminoguanidine. It is recommended
to use a freshly prepared solution of ascorbic
acid to reduce copper (II). THPTA ligand in protein labeling buffer speeds up the reaction by stabilizing
catalytically-active compounds of copper (I). The presence of THPTA ligand also allows protein labeling in water
medium (without organic solvents) and, owing to stabilization of copper (I) oxidation degree, minimizes
production of reactive oxygen species (RAS) and prevents them from damaging proteins by oxidizing
histidine, methionine and cysteine. Aminoguanidine in protein labeling buffer prevents chemically reactive
aldehydes (dehydroascorbate hydrolysis products) from binding to side chains of arginine, N-terminal
cysteine, and lysine.
For this reaction, you will need alkyne- or azide-modified protein in azide
sodium-free buffer, dye azide or alkyne, 1.5х protein labeling buffer, and ascorbic acid. It is recommended to perform
steps 6 to 9 under an inert gas (nitrogen or argon).
We recommend the
following protocol for conjugation of modified proteins with dye derivatives:
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Alkyne derivative of ultra-bright and photostable perylene fluorophore.
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AF 343 (Coumarin) is a blue emitting fluorophore that can serve as a FRET donor for fluorescein (FAM). This azide derivative can be conjugated with alkynes by сlick chemistry. An extended linker arm between the fluorophore and the azide group enhances solubility and provides spatial separation between the dye and the biomolecule.
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PEP (phenylethynylpyrene), polyaromatic hydrocarbon fluorescent dye for click-chemistry labeling.
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Alkyne derivative of yellow-green fluorescent dye AF 430 for copper-catalyzed click reaction with azides.
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Azide derivative of AF 430 for labeling and detecting alkyne-containing biomolecules via сlick reaction.
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Alkyne derivative of AF 488 dye for copper-catalyzed cycloaddition to azides. Pure 5-isomer.
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AF 488 azide is a fluorophore with emission in the green spectrum range. It reacts with alkynes in copper-catalyzed cycloaddition reactions.
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Azide derivative of yellow fluorescent dye AF 555 for сlick сhemistry.
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AF 568 derivative designed for fluorescence microscopy and flow cytometry. Pure 5-isomer.
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Azide derivative of water-soluble AF 568 dye for labeling alkyne-containing biomolecules and fluorescent visualization of biological objects. Pure 6-isomer.
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Azide derivative of water-soluble red-fluorescent dye AF 594 for labeling alkyne-containing biomolecules and fluorescent visualization of biological objects. Pure 6-isomer.
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Bright and photostable azide AF 647 for labeling alkyne-containing biomolecules and fluorescent visualization of biological objects.
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Bifunctional reagent containing alkyne group and hydrazide function. It reacts with aldehydes and ketones. Alkyne group can then be coupled with various azides via CuAAC reaction.
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Maleimide reagent for the attachment of alkyne functional group to proteins, peptides, and other molecules bearing SH (thiol) groups, including cysteine residues.
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Diethyleneglycol (PEG2) based bifunctional linker containing amine and alkyne functional groups.
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Bifunctional diethyleneglycol linker with terminal alkyne and carboxylic acid moieties.
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Bifunctional reactive crosslinker containing terminal alkyne group for click chemistry or Sonogashira coupling, and reactive alkyl iodide group.
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Bifunctional hydrophilic linker with NHS ester and terminal alkyne functional group based on diethyleneglycol (PEG2).
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Alkyne-PEG3-amine is a medium-length heterobifunctional linker for conjugation.
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Bifunctional triethyleneglycol linker with terminal alkyne and carboxylic acid moieties.
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PEG3 with alkyne and iodide functional groups, a reactive and hydrophilic linker for alkylation and click chemistry.
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Bifunctional spacer with alkyne and maleimide groups for conjugation of biomolecules.
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Bifunctional hydrophilic linker with NHS ester and terminal alkyne functional group based on triethyleneglycol (PEG3).
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Alkyne-PEG3-OH is a triethyleneglycol based bifunctional linker with a hydroxyl group and an alkyne fragment.
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A bifunctional linker molecule containing both terminal alkyne moiety for click chemistry, and amine group. Contains a long hydrophilic PEG4 arm.
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Bifunctional tetraethyleneglycol linker with terminal alkyne and carboxylic acid moieties.
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Bifunctional reactive PEG4-based crosslinker containing terminal alkyne group for click chemistry or Sonogashira coupling and reactive alkyl iodide group.
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Water soluble, heterobifunctional hydrophilic linker with a length of 14 Å, containing maleimide for thiol labeling, and alkyne for click chemistry. An ideal reagent to attach an alkyne moiety to a protein.
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Bifunctional hydrophilic linker with NHS ester and terminal alkyne functional group based on tetraethyleneglycol (PEG4).
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Azide derivative of AMCA fluorescent dye for сlick сhemistry. AMCA is one of the brightest blue fluorescent dyes with large Stoke's shift and high resistance to photobleaching.
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Azide derivative of water-soluble yellow-fluorescent dye ATT 550 for labeling alkyne-containing biomolecules and fluorescent visualization of biological objects.
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Azide derivative of water-soluble orange-fluorescent dye ATT 565 for labeling alkyne-containing biomolecules and fluorescent visualization of biological objects. Pure 6-isomer.
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Azide derivative of water-soluble red-fluorescent dye ATT 594 for labeling alkyne-containing biomolecules and fluorescent visualization of biological objects.
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A bifunctional reagent containing the azide group and hydrazide function. It reacts with aldehydes and ketones. The azide group can then be coupled with various alkynes via CuAAC reaction.
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A hydrophilic bifunctional linker with azide and NHS ester function groups flanking PEG6 (hexaethylene glycol).
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Activated NHS ester for azido-labeling of peptides and proteins.
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BDP 558/568 is a borondipyrromethene dye for Cyanine3-channel. This product is a terminal alkyne derivative for copper-catalyzed click chemistry.
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BDP 558/568 is a borondipyrromethene fluorophore with absorption and emission spectra close to Cyanine3. This is an azide derivative for сlick chemistry.
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Azide derivative of BDP 576/589 dye for labeling and detecting alkyne-containing biomolecules via сlick reaction.
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BDP 581/591 is a borondipyrromethene fluorophore useful for fluorescence polarization assays, microscopy, and two-photon experiments. This alkyne is a derivative for copper-catalyzed click chemistry.
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BDP 581/591 is a relatively hydrophobic borondipyrromethene fluorophore that can be used as a probe for ROS species, or fluorescence polarization tracer. This is an azide derivative for сlick chemistry.
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Alkyne dye for copper-catalyzed click chemistry, containing borondipyrromethene fluorophore BDP 630/650 for the red channel.
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BDP 630/650 is a bright borondipyrromethene-based fluorophore for the far red area of the spectrum. It possesses a high extinction coefficient and quantum yield. This is an azide for сlick chemistry labeling.
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BDP 650/665 is a bright borondipyrromethene red-emitting dye. The terminal ethynyl group of BDP 650/665 alkyne can be easily conjugated with various azides using a copper-catalyzed click reaction.
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BDP 650/665 is a red-emitting borondipyrromethene dye. This azide derivative can be conjugated with alkynes and cycloalkynes.
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Borondipyrromethene dye alkyne for click chemistry. Bright and photostable dye for FAM channel.
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BDP FL is a bright and photostable borondipyrromethene dye for the fluorescein channel. This is a dye azide for сlick сhemistry.
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Terminal alkyne derivative of BDP R6G dye for сlick chemistry. BDP R6G is a borondipyrromethene dye whose absorption and emission spectra are similar to R6G (rhodamine 6G).
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BDP R6G is a high performance borondipyrromethene fluorophore which is tuned to match excitation and emission channels of R6G dye.
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Borondipyrromethene dye for TAMRA channel, with alkyne group for copper-catalyzed click chemistry.
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Borondipyrromethene dye with azido group. Spectral properties are similar to TAMRA dye. Due to long fluorescence lifetime, BDP TMR is especially useful for fluorescence polarization measurements.
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BDP TR is a fluorophore for ROX channel. This borondipyrromethene dye is very bright, and resistant to oxidation.
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BDP TR is a red-emitting fluorophore for the ROX channel. This is an azide for copper-catalyzed and copper-free сlick chemistry conjugation.
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Biotin alkyne is a reagent for click chemistry labeling with biotin, an affinity probe to streptavidin.
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Biotin alkyne is a reagent for click chemistry labeling with biotin, an affinity probe to streptavidin.
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Ready-to-use catalytic buffer containing сopper(II) and water-soluble THPTA ligand. It is suitable for click chemistry modification of proteins.
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Cyanine3.5 azide for biomolecule labeling through click chemistry.
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Cyanine3 alkyne is a fluorescent dye alkyne for сlick сhemistry.
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Azide derivative of Cyanine3 fluorescent dye for сlick сhemistry.
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Azide derivative of Cyanine3B fluorescent dye for сlick сhemistry.
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Cyanine5.5 alkyne is a NIR (near infrared) dye azide for copper-catalyzed сlick сhemistry.
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Far red / near infrared fluorescent Cyanine5.5 dye derivative for click chemistry.
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Cyanine5 alkyne is an alkyne dye for click chemistry labeling of biomolecules.
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Cyanine5 azide is a reagent for click chemistry labeling with Cyanine5, a popular fluorescent dye.
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Reactive near infrared (NIR) dye for click chemistry containing a terminal alkyne group.
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Long-wavelength NIR fluorescent dye azide for click chemistry.
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Cyanine7 alkyne is a near infrared (NIR) dye alkyne for click chemistry.
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Near-infrared (NIR) fluorescent dye for click chemistry labeling.
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DusQ 3 dark quencher with azido group for the conjugation with alkyne-containing molecules
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FAM alkyne, 5-isomer - an alkyne derivative of fluorescein dye for click chemistry.
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Fluorescein dye alkyne for click chemistry conjugation, pure 6-isomer.
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FAM (fluorescein) azide for сlick chemistry labeling, pure 5-isomer.
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FAM (fluorescein) azide, 6-isomer is a commonly used fluorescent dye azide for сlick сhemistry.
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Azide of green-yellow emitting HEX dye for conjugation with oligonucleotides via [3+2]-dipolar cycloaddition.
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Hexynoic acid NHS ester for the labeling of biomolecules with alkyne group for copper-catalyzed click chemistry.
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Alkyne activated ester for the modification of amine groups. STP ester group is an excellent alternative to NHS ester functionality.
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JOE dye alkyne derivative for copper-catalyzed click chemistry reaction with azides. JOE dye is widely used for nucleic acid labeling.
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JOE is a chlorinated fluorescein dye which is often used for HEX channel for oligonucleotides. It is less hydrophobic than HEX. This is an azide derivative for сlick chemistry
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An activated ester for the introduction of alkyne into peptides/proteins and click reaction.
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Reagent for fluorescent labeling with pyrene polycyclic aromatic hydrocarbon via click chemistry.
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Pyrene azide with longer triethyleneglycol (PEG3) linker for click chemistry.
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Rhodamine 110 (R110) azide for click chemistry, pure 6-isomer.
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R6G (rhodamine 6G) terminal alkyne for copper-catalyzed click chemistry reaction with azides. Pure 6-isomer of the dye.
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Rhodamine 6G is a xanthene dye that has been used for DNA sequencing and qPCR. This azide derivative is used for the attachment of the dye to DNA and other molecules by means of click chemistry.
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An alkyne derivative of bright red-emitting dye ROX (Rhodamine X, Rhodamine 101) for copper-catalyzed click chemistry bioconjugation. Pure 5-isomer.
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An alkyne derivative of bright red-emitting dye ROX (Rhodamine X, Rhodamine 101) for copper-catalyzed click chemistry bioconjugation. Pure 6-isomer.
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Azide derivative of ROX (Rhodamine X, Rhodamine 101) dye for biomolecule labeling via click chemistry. Pure 5-isomer.
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Azide derivative of ROX (Rhodamine X, Rhodamine 101) dye for biomolecule labeling via click chemistry. Pure 6-isomer.
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Azide of green-yellow emitting dye SIMA for conjugation with oligonucleotides via [3+2]-dipolar cycloaddition.
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Alkyne derivative of water-soluble sulfo-Cyanine3.5 dye for copper-catalyzed click chemistry.
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Azide derivative of water-soluble sulfo-Cyanine3.5 dye for bioconjugation via click chemistry.
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A water-soluble dye alkyne for copper-catalyzed click chemistry. The fluorophore sulfo-Cyanine3 is a bright and photostable dye for the Cy3® channel.
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Water-soluble sulfo-Cyanine3 dye azide for click chemistry labeling of sensitive molecules and intact biological objects.
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Alkyne functionalized, water soluble far red/near infrared emitting fluorophore sulfo-Cyanine5.5.
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sulfo-Cyanine5.5 azide is a derivative of water-soluble far-red emitting fluorescent cyanine dye. The azide group can be conjugated by either copper-catalyzed or copper-free click chemistry with alkynes and cycloalkynes.
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sulfo-Cyanine5 is a water-soluble Cyanine5 dye alkyne for click chemistry.
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Water soluble sulfo-Cyanine5 fluorescent dye azide for click chemistry.
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sulfo-Cyanine7.5 is a near-infrared fluorophore similar to Indocyanine Green (ICG) but possesses higher fluorescence quantum yield. This is an alkyne for copper-catalyzed click chemistry.
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sulfo-Cyanine7.5 is a very hydrophilic and water-soluble dye for the near-infrared region. The absorption and emission spectra of the dye are similar to Indocyanine Green (ICG), but sulfo-Cyanine7.5 has a much higher fluorescence quantum yield. The dye is a great choice for near-infrared in vivo imaging. This is an azide for click chemistry.
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A sulfonated near-infrared dye alkyne for copper-catalyzed cycloaddition with azides.
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Water-soluble NIR fluorescent dye azide for click chemistry.
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Alkyne derivative of TAMRA dye, pure 5-isomer for сopper-catalyzed сlick chemistry.
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Terminal alkyne for copper-catalyzed click chemistry. Tetramethylrhodamine (TAMRA) is a rhodamine dye which is popular in qPCR and other applications. TAMRA is a also a FRET acceptor for FAM.
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Azide derivative of TAMRA fluorescent dye for сlick chemistry.
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Pure 6-isomer of tetramethylrhodamine (TAMRA) dye, azide derivative for copper-catalyzed and copper-free click chemistry reactions with alkynes.
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TET is a green-fluorescent fluorescein derivate with spectral properties similar to R6G, JOE, and VIC, widely used for labeling PCR probes. This reagent is an azide for dye conjugation by click chemistry.
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Azide derivative of red fluorescent dye TR for сlick сhemistry conjugation. Pure 5-isomer.
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VIC is an asymmetric xanthene dye with spectral properties similar to HEX and JOE, widely used for labeling PCR probes. This derivative is an azide for dye conjugation by click chemistry.
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