Click-iT Labeling and Detection Tools
If our ready-to-use kits do not match your experimental requirements, a range of Click-iT tools are available to allow you to develop a labeling and detection strategy that addresses your particular biological questions. In the labeling tools section you will find the primary building blocks for incorporating Click-iT labels into the proteins or nucleic acids in your biological model. The table of detection tools shows the complementary probes offered with fluorescent tags that provide visualization or capture of your target molecule.
On this page:
Labeling tools
Protein and glycan labeling tools
We offer Click-iT tools for labeling nascent protein synthesis and for detecting a range of post-translational modifications including fatty acid attachment and glycosylation.
Nucleic acid labeling tools
Click chemistry tools can be used to either label or capture nascent DNA or RNA for subsequent detection or analysis.
Target | Product | Tag | Quantity | Cat. No. |
---|---|---|---|---|
Nascent Proteins | Click-iT AHA (L-Azidohomoalanine) | azide | 5 mg | C10102 |
Click-iT HPG (L-homopropargylglycine) | alkyne | 5 mg | C10186 | |
OPP (O-propargyl-puromycin) | alkyne | 5mg | C10459 | |
Farnesylated proteins | Click-iT farnesyl alcohol, azide | azide | 5 mg | C10248 |
Geranylgeranylated proteins | Click-iT geranylgeranyl alcohol | azide | 5 mg | C10249 |
Fucosylated glycans | Click-iT fucose alkyne | alkyne | 5 mg | C10264 |
Palmitylated proteins | Click-iT palmitic acid, azide | azide | 1 mg | C10265 |
Myristoylated proteins | Click-iT myristic acid, azide | azide | 1 mg | C10268 |
O-Linked glycoproteins | Click-iT GalNAz (tetraacetylated N-azidoacetylgalactosamine) | azide | 1 each | C33365 |
Sialic acid-modified glycoproteins | Click-iT ManNAz (tetraacetylated N-azidoacetyl-D-mannosamine | azide | 1 each | C33366 |
O-GlcNAz-modified glycoproteins | Click-iT GlcNAz (tetraacetylated N-azidoacetylglucosamine) | azide | 1 each | C33367 |
Target | Product | Tag | Quantity | Cat. No. |
---|---|---|---|---|
Nascent DNA | EdU (5-ethynyl-2′-deoxyuridine) | alkyne | 50 mg | A10044 |
EdU (5-ethynyl-2′-deoxyuridine) | alkyne | 500 mg | E10187 | |
Nascent RNA | 5-Ethynyl Uridine (EU) | alkyne | 5 mg | E10345 |
Target | Product | Tag | Quantity | Cat. No. |
---|---|---|---|---|
Nascent Proteins | Click-iT AHA (L-Azidohomoalanine) | azide | 5 mg | C10102 |
Click-iT HPG (L-homopropargylglycine) | alkyne | 5 mg | C10186 | |
OPP (O-propargyl-puromycin) | alkyne | 5mg | C10459 | |
Farnesylated proteins | Click-iT farnesyl alcohol, azide | azide | 5 mg | C10248 |
Geranylgeranylated proteins | Click-iT geranylgeranyl alcohol | azide | 5 mg | C10249 |
Fucosylated glycans | Click-iT fucose alkyne | alkyne | 5 mg | C10264 |
Palmitylated proteins | Click-iT palmitic acid, azide | azide | 1 mg | C10265 |
Myristoylated proteins | Click-iT myristic acid, azide | azide | 1 mg | C10268 |
O-Linked glycoproteins | Click-iT GalNAz (tetraacetylated N-azidoacetylgalactosamine) | azide | 1 each | C33365 |
Sialic acid-modified glycoproteins | Click-iT ManNAz (tetraacetylated N-azidoacetyl-D-mannosamine | azide | 1 each | C33366 |
O-GlcNAz-modified glycoproteins | Click-iT GlcNAz (tetraacetylated N-azidoacetylglucosamine) | azide | 1 each | C33367 |
Target | Product | Tag | Quantity | Cat. No. |
---|---|---|---|---|
Nascent DNA | EdU (5-ethynyl-2′-deoxyuridine) | alkyne | 50 mg | A10044 |
EdU (5-ethynyl-2′-deoxyuridine) | alkyne | 500 mg | E10187 | |
Nascent RNA | 5-Ethynyl Uridine (EU) | alkyne | 5 mg | E10345 |
Detection tools
Standard reactions
Azides and alkynes can be used interchangeably in standard click reactions, so either one can serve as the tagged substrate or be labeled for the detection step. Some biomolecules are sensitive to the amount of copper used to catalyze standard click reactions. In particular, proteins such as GFP and R-PE exhibit reduced fluorescence and nucleic acids can be denatured by a high-copper environment.
Copper sensitivity
Biomolecules exhibit varying sensitivity to copper concentration. The high copper concentration used to catalyze traditional click reactions maintains the fluorescence of APC, but not GFP. At medium copper concentrations, GFP and APC fluorescence is maintained, but phalloidin binding is lost (Table 1). With Click-iT Plus reagents, the copper concentration can be optimized to preserve the signal in your fluorescent proteins and also drive the EdU click reaction.
Table 1. The effects of copper on fluorescence and phalloidin binding.
Copper concentration | Maintained | Lost |
---|---|---|
High |
|
|
Medium |
|
|
Low |
|
Picolyl azides for low-copper reactions
Click-iT Plus reagents enable researchers to perform copper-catalyzed click reactions with compounds that are sensitive to copper, while retaining all of the benefits of the original azide/alkyne click reaction. Each Click-iT Plus picolyl azide–based tool box includes everything required to optimize the copper concentration and protect against undesired copper side reactions with proteins (e.g., GFP, RPE), nucleic acids (e.g., RNA, oligos), and even small molecules (e.g., phalloidin).
Additionally the Click-iT Plus picolyl azides can be used to increase sensitivity or accelerate the reactions by using the original copper-catalyzed reaction conditions.
DIBO alkynes for copper-free reactions
DIBO reagents can be used to perform click reactions with azide-modified targets without the use of heavy metal catalysis. DIBO reactions are ideal for surface labeling of live cells and also minimize damage to fluorescent proteins like GFP or R-PE. For intracellular labeling, the nonspecific binding of DIBO alkynes creates a background signal, and a labeling strategy using picolyl azides is the preferred solution in these instances.
Label | Product | Tag | Quantity | Cat. No. |
---|---|---|---|---|
Alexa Fluor 488 | Click-iT Plus Alexa Fluor 488 picolyl azide toolkit (includes reaction buffers) | picolyl azide | 1 kit, 30 to 50 reactions depending on application | C10641 |
Alexa Fluor 555 | Click-iT Plus Alexa Fluor 555 picolyl azide toolkit (includes reaction buffers) | picolyl azide | 1 kit, 30 to 50 reactions depending on application | C10642 |
Alexa Fluor 647 | Click-iT Plus Alexa Fluor 647 picolyl azide toolkit (includes reaction buffers) | picolyl azide | 1 kit, 30 to 50 reactions depending on application | C10643 |
Label | Product | Tag | Quantity | Cat. No. |
---|---|---|---|---|
Alexa Fluor 488 | Click-iT Alexa Fluor 488 DIBO alkyne | alkyne | 0.5 mg | C10405 |
Alexa Fluor 555 | Click-iT Alexa Fluor 555 DIBO alkyne | alkyne | 0.5 mg | C10406 |
Alexa Fluor 594 | Click-iT Alexa Fluor 594 DIBO alkyne | alkyne | 0.5 mg | C10407 |
Alexa Fluor 647 | Click-iT Alexa Fluor 647 DIBO alkyne | alkyne | 0.5 mg | C10408 |
TAMRA/tetramethylrhodamine | Click-iT TAMRA DIBO alkyne | alkyne | 0.5 mg | C10410 |
Biotin | Click-iT biotin DIBO alkyne | alkyne | 1 mg | C10412 |
No label, reactive (amine containing) | Click-iT amine DIBO alkyne | alkyne | 1 mg | C10411 |
No label, thiol reactive | Click-iT maleimide DIBO alkyne | alkyne | 1 mg | C10413 |
No label, amine reactive NHS ester | Click-iT succinimidyl ester DIBO alkyne | alkyne | 1 mg | C10414 |
Label | Product | Tag | Quantity | Cat. No. |
---|---|---|---|---|
Alexa Fluor 488 | Click-iT Plus Alexa Fluor 488 picolyl azide toolkit (includes reaction buffers) | picolyl azide | 1 kit, 30 to 50 reactions depending on application | C10641 |
Alexa Fluor 555 | Click-iT Plus Alexa Fluor 555 picolyl azide toolkit (includes reaction buffers) | picolyl azide | 1 kit, 30 to 50 reactions depending on application | C10642 |
Alexa Fluor 647 | Click-iT Plus Alexa Fluor 647 picolyl azide toolkit (includes reaction buffers) | picolyl azide | 1 kit, 30 to 50 reactions depending on application | C10643 |
Label | Product | Tag | Quantity | Cat. No. |
---|---|---|---|---|
Alexa Fluor 488 | Click-iT Alexa Fluor 488 DIBO alkyne | alkyne | 0.5 mg | C10405 |
Alexa Fluor 555 | Click-iT Alexa Fluor 555 DIBO alkyne | alkyne | 0.5 mg | C10406 |
Alexa Fluor 594 | Click-iT Alexa Fluor 594 DIBO alkyne | alkyne | 0.5 mg | C10407 |
Alexa Fluor 647 | Click-iT Alexa Fluor 647 DIBO alkyne | alkyne | 0.5 mg | C10408 |
TAMRA/tetramethylrhodamine | Click-iT TAMRA DIBO alkyne | alkyne | 0.5 mg | C10410 |
Biotin | Click-iT biotin DIBO alkyne | alkyne | 1 mg | C10412 |
No label, reactive (amine containing) | Click-iT amine DIBO alkyne | alkyne | 1 mg | C10411 |
No label, thiol reactive | Click-iT maleimide DIBO alkyne | alkyne | 1 mg | C10413 |
No label, amine reactive NHS ester | Click-iT succinimidyl ester DIBO alkyne | alkyne | 1 mg | C10414 |
Buffers for standard Click-iT reactions
For added convenience, we offer Click-iT Reaction Buffer Kits optimized for either proteins or cell samples labeled with an azide- or alkyne-tagged biomolecule.
- The Click-iT Cell Reaction Buffer Kit includes sufficient reagents to perform 50 reactions based on a 0.5 mL reaction volume for subsequent analyses by flow cytometry, fluorescence microscopy, or high-content screening (HCS).
- The Click-iT Protein Reaction Buffer Kit includes everything required to perform the click reaction of proteins for subsequent standard protein biochemical analyses (e.g., western blots, mass spectrometry).
Resources
Learn more
Molecular Probes Handbook
BioProbes Journal articles
Scientific posters
For Research Use Only. Not for use in diagnostic procedures.