Click Chemistry Tools for Biological Assays
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Click Chemistry—Making Biodiscovery a Snap
Click Chemistry
Click chemistry describes a powerful new class of chemical reactions that fill the void left when direct labeling or antibodies fail or simply fall short. Click reactions use biologically unique and inert moieties to label and eventually detect a molecule using a two-step procedure.
Click Reaction in Biomedical Research
Life Technologies is pleased to offer the largest selection of assays and reagents based upon the click reaction involving the copper-catalyzed triazole formation of an azide and an alkyne, the most widely used click reaction in biomedical research.
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Figure 1. Click azide/alkyne reaction. The azide and alkyne moieties are interchangeable, whereupon the molecule can be labeled with an alkyne or azide and be detected with the corresponding a fluorophore- or hapten-azide or -alkyne.
Small Size Translates to Big Advantages
Azides/Alkynes are Unique, Inert and Stable
Both azides and alkynes are biologically unique, inert, stable—and extremely small (Figure 2). These moieties are interchangeable; either one can tag your target molecule, while the other is used for subsequent detection. The label is small enough that tagged molecules are acceptable substrates for enzymes that assemble biopolymers.
Both azides and alkynes are biologically unique, inert, stable—and extremely small (Figure 2). These moieties are interchangeable; either one can tag your target molecule, while the other is used for subsequent detection. The label is small enough that tagged molecules are acceptable substrates for enzymes that assemble biopolymers.
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Figure 2. Relative size of detection molecules commonly used in cellular analysis.
Table 1. Click-iT® Metabolic Labeling Reagents for Proteins
| Application | Compound | Azide or alkyne | Cat. no. |
|---|---|---|---|
| Monitor nascent protein synthesis or inhibition | Click-iT® AHA (L-azidohomoalanine) | azide | C10102 |
| Click-iT® HPG (L-homopropargylglycine) | alkyne | C10186 | |
| Identification of isoprenylated proteins | Click-iT® farnesyl alcohol, azide | azide | C10248 |
| Click-iT® geranylgeranyl alcohol, azide | azide | C10249 | |
| Identification of fucoslyated proteins | Click-iT® fucose alkyne (tetraacetylfucose alkyne) | alkyne | C10264 |
| Identification of protein fatty acylation | Click-iT® palmitic acid, azide | azide | C10265 |
| Click-iT® myristic acid, azide | azide | C10268 | |
| Identification of O-linked glycoproteins, including O-GlcNAc | Click-iT® GalNAz (tetraacetylated N-azidoacetylgalactosamine) | azide | C33365 |
| Identification of sialic acid-modified glycoproteins | Click-iT® ManNAz (tetraacetylated N-azidoacetylmannosamine) | azide | C33366 |
| Identification of O-GlcNAc-modified glycoproteins | Click-iT® GlcNAz (tetraacetylated N-azidoacetylglucoosamine) | azide | C33367 |
Table 2. Azide-or Alkyne-Modified Fluorophores and Haptens
| Label | Ex/Em* | Azide or alkyne | Cat. no. | Use |
|---|---|---|---|---|
| Alexa Fluor® 488 | 495/519 | azide | A10266 | Fluorescent dye or hapten** |
| alkyne | A10267 | |||
| Alexa Fluor® 555 | 555/565 | azide | A20012 | Fluorescent dye |
| alkyne | A20013 | |||
| Alexa Fluor® 594 | 590/617 | azide | A10270 | Fluorescent dye |
| alkyne | A10275 | |||
| Alexa Fluor® 647 | 650/668 | azide | A10277 | Fluorescent dye |
| alkyne | A10278 | |||
| Biotin † | NA | azide | B10184 | Hapten |
| alkyne | B10185 | |||
| Oregon Green® 488 | 496/524 | azide | O10180 | Fluorescent dye or hapten** |
| alkyne | O10181 | |||
| Tetramethylrhodamine (TAMRA) | 555/580 | azide | T10182 | Fluorescent dye or hapten** |
| alkyne | T10183 | |||
| Click-iT® Alexa Fluor® 488 DIBO | 495/519 | alkyne | C10405 | Copper free click chemistry detection of azide |
| Click-iT® Alexa Fluor® 555 DIBO | 555/565 | alkyne | C10406 | Copper free click chemistry detection of azide |
| Click-iT® Alexa Fluor® 594 DIBO | 590/617 | alkyne | C10407 | Copper free click chemistry detection of azide |
| Click-iT® Alexa Fluor® 647 DIBO | 650/668 | alkyne | C10408 | Copper free click chemistry detection of azide |
| Click-iT® TAMRA DIBO | 542/568 | alkyne | C10410 | Copper free click chemistry detection of azide |
| Click-iT® amine DIBO | To create amine-modified reagents | alkyne | C10411 | Copper free click chemistry detection of azide |
| Click-iT® biotin DIBO | Hapten (biotin) | alkyne | C10412 | Copper free click chemistry detection of azide |
| Click-iT® maleimide DIBO | Thiol Reactive Form | alkyne | C10413 | Copper free click chemistry detection of azide |
| Click-iT® succinimidyl ester DIBO | Amine Reactive Form | alkyne | C10414 | Copper free click chemistry detection of azide |
| *Absorption and fluorescence emission maxima, in nm. † Requires streptavidin for detection. **As hapten, use with anti-dye antibody. | ||||
Performing Click-iT® Reactions
For added convenience, Life Technologies offers Click-iT® Reaction Buffer Kits for protein 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).
Find Your Click-iT® Detection Assay
With its equally small “footprint,” the Click-iT® detection molecule can easily penetrate complex samples for in-depth analysis of key biological processes, including assays for:
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Cutting-edge Click-iT® Citations
- EdU, a new Thymidine Analogue for Labeling Proliferating Cells in the Nervous System—J. Neurosci Meth. 177, 122-130 (2009)
- Exploring RNA Transcription and turnover in-vivo by using Click Chemistry—PNAS 105, 15779-15784 (2008)
