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

Protein and glycan labeling tools

Life Technologies offers 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.

 

Click-iT® Labeling Tools
Proliferating cells in rat ileum using Click-iT® EdU.

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
  • 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.

 

Label Product Tag Quantity Cat. No.
Alexa Fluor®488 Alexa Fluor® 488 azide, triethylammonium salt azide 0.5 mg A10266
Alexa Fluor® 555 Alexa Fluor® 555 azide, triethylammonium salt azide 0.5 mg A20012
Alexa Fluor® 594 Alexa Fluor® 594 azide, triethyammonium salt azide 0.5 mg A10270
Alexa Fluor® 647 Alexa Fluor® 647 azide, triethylammonium salt azide 0.5 mg A10277
Oregon Green® 488 azide Oregon Green® 488 azide®, triethylammonium salt azide 0.5 mg O10180
TAMRA Tetramethylrhodamine (TAMRA) azide azide 0.5 mg T10182
Biotin Biotin azide azide 1 mg B10184
Amine reactive (NHS ester) Azido (PEO)4 propionic acid, succinimidyl ester azide 1 mg A10280
No label, thiol reactive Iodoacetamide azide azide 1 mg I10188
Alexa Fluor® 488 Alexa Fluor® 488 alkyne, triethylammonium salt alkyne 0.5 mg A10267
Alexa Fluor® 555 Alexa Fluor® 555 alkyne, triethylammonium salt alkyne 0.5 mg A20013
Alexa Fluor® 594 Alexa Fluor® 594 alkyne, triethylammonium salt alkyne 0.5 mg A10275
Alexa Fluor® 647 Alexa Fluor® 647 alkyne, triethylammonium salt alkyne 0.5 mg A10278
TAMRA/ tetramethylrhodamine Tetramethylrhodamine (TAMRA) alkyne alkyne 0.5 mg T10183
Biotin Biotin alkyne alkyne 1 mg B10185
No label, amine reactive (NHS ester) Alkyne, succinimidyl ester alkyne 1 mg A10279
No label, thiol reactive Iodoacetamide alkyne alkyne 1 mg I10189
* Optimized buffers are not supplied with these reagents and are available for purchase separately.
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).