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.

Labeling tools

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.

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 concentrationMaintainedLost
High
  • APC fluorescence
  • GFP fluorescence
  • Phalloidin binding
Medium
  • APC fluorescence
  • GFP fluorescence
  • Phalloidin binding
Low
  • APC fluorescence
  • GFP fluorescence
  • Phalloidin binding
 

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.

LabelProductTagQuantityCat. No.
Alexa Fluor 488Alexa Fluor 488 azide, triethylammonium salt azide0.5 mgA10266
Alexa Fluor 555Alexa Fluor 555 azide, triethylammonium salt azide0.5 mgA20012
Alexa Fluor 594Alexa Fluor 594 azide, triethylammonium salt azide0.5 mgA10270
Alexa Fluor 647Alexa Fluor 647 azide, triethylammonium salt azide0.5 mgA10277
Oregon Green 488 azideOregon Green 488 azide, triethylammonium saltazide0.5 mgO10180
TAMRATetramethylrhodamine (TAMRA) azide azide0.5 mgT10182
BiotinBiotin azideazide1 mgB10184
Amine reactive (NHS ester)Azido (PEO)4 propionic acid, succinimidyl esterazide1 mgA10280
No label, thiol reactiveIodoacetamide azideazide1 mgI10188
Alexa Fluor 488Alexa Fluor 488 alkyne, triethylammonium saltalkyne0.5 mgA10267
Alexa Fluor 555Alexa Fluor 555 alkyne, triethylammonium salt alkyne0.5 mgA20013
Alexa Fluor 594Alexa Fluor 594 alkyne, triethylammonium saltalkyne0.5 mgA10275
Alexa Fluor 647Alexa Fluor 647 alkyne, triethylammonium salt alkyne0.5 mgA10278
TAMRA/ tetramethylrhodamineTetramethylrhodamine (TAMRA) alkynealkyne0.5 mgT10183
BiotinBiotin alkyne alkyne1 mgB10185
No label, amine reactive (NHS ester)Alkyne, succinimidyl esteralkyne1 mgA10279
No label, thiol reactiveIodoacetamide alkyne alkyne1 mgI10189
* Optimized buffers are not supplied with these reagents and are available for purchase separately.
LabelProductTagQuantityCat. No.
Alexa Fluor 488Click-iT Plus Alexa Fluor 488 picolyl azide toolkit (includes reaction buffers)picolyl azide1 kit, 30 to 50 reactions depending on applicationC10641
Alexa Fluor 555Click-iT Plus Alexa Fluor 555 picolyl azide toolkit (includes reaction buffers)picolyl azide1 kit, 30 to 50 reactions depending on applicationC10642
Alexa Fluor 647Click-iT Plus Alexa Fluor 647 picolyl azide toolkit (includes reaction buffers)picolyl azide1 kit, 30 to 50 reactions depending on applicationC10643
LabelProductTagQuantityCat. No.
Alexa Fluor 488Click-iT Alexa Fluor 488 DIBO alkynealkyne0.5 mgC10405
Alexa Fluor 555Click-iT Alexa Fluor 555 DIBO alkynealkyne0.5 mgC10406
Alexa Fluor 594Click-iT Alexa Fluor 594 DIBO alkynealkyne0.5 mgC10407
Alexa Fluor 647Click-iT Alexa Fluor 647 DIBO alkynealkyne0.5 mgC10408
TAMRA/tetramethylrhodamineClick-iT TAMRA DIBO alkynealkyne0.5 mgC10410
BiotinClick-iT biotin DIBO alkynealkyne1 mgC10412
No label, reactive (amine containing)Click-iT amine DIBO alkynealkyne1 mgC10411
No label, thiol reactiveClick-iT maleimide DIBO alkynealkyne1 mgC10413
No label, amine reactive NHS ester Click-iT succinimidyl ester DIBO alkynealkyne1 mgC10414
LabelProductTagQuantityCat. No.
Alexa Fluor 488Alexa Fluor 488 azide, triethylammonium salt azide0.5 mgA10266
Alexa Fluor 555Alexa Fluor 555 azide, triethylammonium salt azide0.5 mgA20012
Alexa Fluor 594Alexa Fluor 594 azide, triethylammonium salt azide0.5 mgA10270
Alexa Fluor 647Alexa Fluor 647 azide, triethylammonium salt azide0.5 mgA10277
Oregon Green 488 azideOregon Green 488 azide, triethylammonium saltazide0.5 mgO10180
TAMRATetramethylrhodamine (TAMRA) azide azide0.5 mgT10182
BiotinBiotin azideazide1 mgB10184
Amine reactive (NHS ester)Azido (PEO)4 propionic acid, succinimidyl esterazide1 mgA10280
No label, thiol reactiveIodoacetamide azideazide1 mgI10188
Alexa Fluor 488Alexa Fluor 488 alkyne, triethylammonium saltalkyne0.5 mgA10267
Alexa Fluor 555Alexa Fluor 555 alkyne, triethylammonium salt alkyne0.5 mgA20013
Alexa Fluor 594Alexa Fluor 594 alkyne, triethylammonium saltalkyne0.5 mgA10275
Alexa Fluor 647Alexa Fluor 647 alkyne, triethylammonium salt alkyne0.5 mgA10278
TAMRA/ tetramethylrhodamineTetramethylrhodamine (TAMRA) alkynealkyne0.5 mgT10183
BiotinBiotin alkyne alkyne1 mgB10185
No label, amine reactive (NHS ester)Alkyne, succinimidyl esteralkyne1 mgA10279
No label, thiol reactiveIodoacetamide alkyne alkyne1 mgI10189
* Optimized buffers are not supplied with these reagents and are available for purchase separately.
LabelProductTagQuantityCat. No.
Alexa Fluor 488Click-iT Plus Alexa Fluor 488 picolyl azide toolkit (includes reaction buffers)picolyl azide1 kit, 30 to 50 reactions depending on applicationC10641
Alexa Fluor 555Click-iT Plus Alexa Fluor 555 picolyl azide toolkit (includes reaction buffers)picolyl azide1 kit, 30 to 50 reactions depending on applicationC10642
Alexa Fluor 647Click-iT Plus Alexa Fluor 647 picolyl azide toolkit (includes reaction buffers)picolyl azide1 kit, 30 to 50 reactions depending on applicationC10643
LabelProductTagQuantityCat. No.
Alexa Fluor 488Click-iT Alexa Fluor 488 DIBO alkynealkyne0.5 mgC10405
Alexa Fluor 555Click-iT Alexa Fluor 555 DIBO alkynealkyne0.5 mgC10406
Alexa Fluor 594Click-iT Alexa Fluor 594 DIBO alkynealkyne0.5 mgC10407
Alexa Fluor 647Click-iT Alexa Fluor 647 DIBO alkynealkyne0.5 mgC10408
TAMRA/tetramethylrhodamineClick-iT TAMRA DIBO alkynealkyne0.5 mgC10410
BiotinClick-iT biotin DIBO alkynealkyne1 mgC10412
No label, reactive (amine containing)Click-iT amine DIBO alkynealkyne1 mgC10411
No label, thiol reactiveClick-iT maleimide DIBO alkynealkyne1 mgC10413
No label, amine reactive NHS ester Click-iT succinimidyl ester DIBO alkynealkyne1 mgC10414

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).