2-Ethynyl-ATP (2-EATP)

Product Information

Alkyne‑Modified ATP for Click‑Enabled RNA Labeling & ATP‑Dependent Enzyme Studies

2‑Ethynyl‑ATP (2‑EATP) is a terminal‑alkyne–modified adenosine‑5′-triphosphate designed for biochemical assays, RNA labeling workflows, and bioorthogonal click chemistry. Its reactive ethynyl group enables highly efficient copper(I)‑catalyzed azide‑alkyne cycloaddition (CuAAC), allowing fluorescent dyes, biotin labels, affinity handles, or biomolecules to be covalently attached.

2‑EATP is widely used in ATP‑binding studies, ATP‑dependent enzyme assays, in vitro RNA polyadenylation, and advanced RNA imaging and quantification techniques.

Key Features

• Alkyne-functionalized ATP analog for CuAAC click chemistry
• Suitable for ATP-binding assays, kinase studies, ATPase assays, and enzyme kinetics
• Efficiently incorporated by recombinant poly(A) polymerase for RNA 3’-end labeling
• Enables flexible downstream modification with:
  • Biotin‑azide
  • Fluorophore‑azides (488, 555, 594, 647)
  • Azide‑linked crosslinkers
• High purity (≥ 90%, HPLC)
• Supplied as a ready‑to‑use 10 mM solution
• Long shelf life (12 months at –20 °C, dry, dark)

How 2‑Ethynyl‑ATP Works

1. Enzymatic Incorporation: 2‑EATP is recognized by several ATP‑dependent enzymes including:

• • Recombinant poly(A) polymerase
  • Kinases
  • ATPases
  • RNA‑processing enzymes

This enables biochemical characterization of ATP‑dependent mechanisms.

2. RNA Polyadenylation and 3’-End Labeling

During in vitro polyadenylation, 2‑EATP is incorporated into nascent poly(A) tails, generating click‑reactive RNA molecules suitable for downstream functionalization.

3. CuAAC Click Chemistry Functionalization

The terminal alkyne reacts with azides to attach:

• • Biotin (for purification workflows)
  • Fluorophores (for RNA imaging)
  • Protein conjugates (for crosslinking analyses)

Applications

1. ATP‑Dependent Enzyme Studies

Ideal for investigating:

• • ATP‑binding proteins
  • Kinases
  • ATPases
  • Enzymatic mechanisms relevant to cancer and neurological disorders

2. In Vitro Polyadenylation of RNA

Enables:

• • Functionalized RNA 3’-end labeling
  • Poly(A)-profiling
  • RNA Tail engineering for mechanistic studies

3. Click‑Enabled RNA Labeling and Purification

Allows downstream introduction of:

• • Biotin tags
  • Fluorescent dyes
  • Bioorthogonal linkers
  • for imaging, pull‑down assays, or conjugations.

4. Advanced RNA Imaging & Analysis Technologies

Used in high‑impact methods such as:

• • Rolling FISH (rFISH)
  • Single‑cell polyadenylation profiling
  • Poly(A)-trap assays

LITERATURE

Click-encoded rolling FISH for visualizing single-cell RNA polyadenylation and structures, F. Chen et al., 2019, Nucleic Acids Research, Volume 47, Issue 22, Page e145.

https://doi.org/10.1093/nar/gkz852

Global profiling of stimulus-induced polyadenylation in cells using a poly(A) trap, D. Curanovic et al., 2013, Nat Chem Biol, Vol. 9, p. 671–673.

https://doi.org/10.1038/nchembio.1334

Template-independent enzymatic synthesis of RNA oligonucleotides, D. J. Wiegand et al., 2025, Nature Biotechnology, Vol. 43(5), p. 762–772.

https://doi.org/10.1038/s41587-024-02244-w

Click-encoded rolling FISH for visualizing single-cell RNA polyadenylation and structures, F. Chen et al., 2019, Nucleic Acids Research, Vol. 47(22), p. e145.

https://doi.org/10.1093/nar/gkz852

Global profiling of stimulus-induced polyadenylation in cells using a poly(A) trap, D. Curanovic et al., 2013, Nature Chemical Biology, Vol. 9, p. 671–673.

https://doi.org/10.1038/nchembio.1334

A Platform for Controlled Template-Independent Enzymatic Synthesis of RNA Oligonucleotides and Therapeutics, D. J. Wiegand et al., 2023, bioRxiv, 2023-06.

https://doi.org/10.1101/2023.06.29.547106