Quenched Fluorescent Peptide (FRET Peptide) Synthesis

Quenched fluorescent provides a safer alternative to the use of radiolabeled isotopes. It's rapid, sensitive, and easily automated. CD Formulation emerges as a vanguard in the realm of quenched fluorescent peptide (FRET peptide) technology. Our custom services in FRET peptide synthesis cater to an eclectic tapestry of needs—be it the meticulous crafting of a solitary FRET peptide or the adventurous endeavor of orchestrating an extensive FRET peptide library.

What Are FRET Peptides

Quenched fluorescent peptides, often referred to as "FRET peptides", emerge as captivating instruments in the labyrinth of biochemical exploration, illuminating the mystifying choreography of enzyme dynamics, untangling the intricate tapestry of protein-protein interactions, and navigating the convoluted routes of subcellular protein transport—among myriad other vibrant biological functions. These remarkable peptides are synthesized through the intricate fusion of synthetic protein fragments with fluorophore and quencher dyes.

Fig. 1 Schematic diagram of FRET peptide mechanism. (CD Formulation)

Explore Our Custom FRET Peptide Synthesis Services

CD Formulation has extensive knowledge in FRET peptide design and synthesis. We offer a variety of FRET substrates to meet your research needs, including pre-made FRET peptides or custom FRET sequences.

Standard dye combinations for FRET:

• Abz (2-Aminobenzoyl) / Dnp (2,4-Dinitrophenyl)
• EDANS (5-[(2-Aminoethyl) amino] naphthalene-1-sulfonic acid) / Dabcyl (4-(4-Dimethylaminophenylazo)benzoyl)
• Methoxycoumarin acetic acid (MCA) and 2,4-dinitrophenyl (DNP)

The Abz/Dnp pair has the highest success rate in peptide synthesis. Typically, the Dnp quencher is conjugated internally or at the C-terminus to the primary amino group on a lysine residue, while the fluorescent dye is Abz conjugated to the N-terminus.

The EDANS/Dabcyl pair is an alternative to the Abz/Dnp pair. However, this pair is significantly more expensive to synthesize and more complex to incorporate during peptide synthesis than the Abz/Dnp pair.

Below, we provide a table of commonly used fluorophore/quencher pairs:

Design Your FRET Peptide

Standard FRET peptides are labeled with a donor/fluorescent molecule and an acceptor/quencher molecule. CD Formulation's FRET peptides are carefully designed to ensure accurate attachment of the fluorophore and quencher. The most common attachment options include N-terminal and C-terminal conjugation, ensuring flexibility in peptide design. These methods allow for the customization of FRET probes to the specific needs of a particular experiment.

Here are the considerations we take into account when designing and selecting FRET pairs:

• Amino acid sequence: The efficiency of energy transfer depends on distance, and our scientists are able to design the most appropriate amino acid sequence length, as the length of the peptide is critical for the correct FRET signal generation.
• Spectral compatibility: Our team of brilliant scientists stands ready to assist you in the intricate selection of FRET pairs, artfully navigating the labyrinth of spectral characteristics. By deftly minimizing the overlap between excitation and emission spectra, we aim to elevate your detection accuracy to unprecedented heights, ensuring that every subtle nuance is captured with precision.
• Quenchers: Certain quenchers, like Dabcyl and Dnp, wield remarkable prowess in obliterating background fluorescence, thus rendering them particularly adept for high-sensitivity assays. In the labyrinth of research, our scientists meticulously sift through an array of fluorophore and quencher pairings, diligently curating the optimal selections tailored to your specific investigative pursuits.

TR-FRET Peptide

Time-resolved FRET (TR-FRET) is a method that utilizes long-lived fluorophores (characteristic of lanthanides) to delay the measurement by 50–150 µs.

• In TR-FRET, lanthanide metals such as europium (Eu) are used as donor molecules instead of fluorescent dyes. This europium fluorescence has a longer lifetime than traditional fluorophores. Due to the large Stokes shift of europium (Eu), TR-FRET provides high signal-to-noise ratios with minimal crosstalk between excitation and emission wavelengths. TR-FRET allows for flexible, reliable, and sensitive detection with fewer false results in high-throughput applications.
• DOTA is a metal chelator that binds to lanthanide metals, and we can synthesize any DOTA peptide for any TR-FRET application.

Applications of FRET Peptide

• In vitro and in vivo subcellular imaging.
• Biosensing.
• Protein-protein interaction studies.
• Enzyme research.

Peptide Manufacturing & Analytical Services

In addition to peptide synthesis capabilities, CD Formulation combines flexible GMP manufacturing facilities with cutting-edge peptide analytical knowledge to provide a full range of quality control testing services to accelerate the commercialization of your products, including:

Publication

CD Formulation is a trusted partner for peptide synthesis. Please don't hesitate to contact us if you are considering using FRET peptides in your project. We look forward to cooperating with you.

How It Works

STEP 1

Submit a service request describing your specific research or development need.

STEP 2

We'll email you to provide your quote and confirm order details if applicable.

STEP 3

Execute the project with real-time communication, and deliver the final report promptly.