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    ADME-Tox
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    Drug Discovery
    Gene Expression - Molecular Biology
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Tebubio's blog - Acting and reacting in life sciences and biotechnologies
  • Home
  • Research areas
    • ADME-Tox
    • Biomarkers
    • Cell Biology and Signalling
    • Cell Sourcing – Cell Culture Technologies
    • Drug Discovery
    • Gene Expression – Molecular Biology
    • Stem Cells
    • Supplying Discovery Tools
  • Contact us
  • Meet the authors
Supplying Discovery Tools

How can TR-FRET facilitate your cellular studies?

06/01/2020 by Frédéric Samazan No Comments
Inflammatory Cytokine Array and ELISA Services tebu-bio

Time-Resolved Förster Resonance Energy Transfer (TR-FRET) is a robust and homogeneous fluorescence detection technology and a real alternative to classical ELISA and Western Blot to perform Cell Signalling studies. Based on this improved TR-FRET technology, tebu-bio is pleased to introduce a new product line of high-quality, fully-validated and affordable kits. These ready-to-use Thunder™ kits enable sensitive, simple and rapid measurement of low amounts of specific intracellular phosphorylated and total proteins in cell lysates from adherent or suspension cells.

The main features of these new Thunder™ TR-FRET kits are presented in this post.

How does it work?

Even though based on the traditional immuno-assay sandwich principle, the kits use a streamlined protocol wherein the antibody-target sandwich complex is formed in solution in a single addition and incubation step, without any wash steps. This protocol dramatically decreases hands-on time (5 minutes) and enables faster time to results (1-4 hours).

Figure 1:TR-FRET principle

After lysis steps, a pair of antibodies specific for the target protein is added to the lysate sample (Fig.1). One antibody is labeled with a donor fluorophore (a Europium chelate), while the second antibody is labeled with an acceptor fluorophore (a far-red dye). Upon excitation of the Europium chelate at 320 or 340 nm, energy is transferred from the donor to the acceptor fluorophore if they are sufficiently close (within 10 nm) for FRET. This results in the emission by the acceptor of a TR-FRET signal at 665 nm.

Why is THUNDER TR-FRET a smart solution for cellular studies?

OPTIMAL ANTIBODY PAIR SELECTION

  • Validated antibodies were selected to guarantee specificity for the target protein.
  • Many donor/acceptor fluorophores were tested to select the optimal pair providing the best TR-FRET signal-to-background.
  • Extensive testing of labeled antibody pairs using optimized lysis buffers followed by assay optimization were conducted to ensure specificity, reproducibility and optimal TR-FRET assay performance.

RIGOROUS VALIDATION

All Thunder™ kits are subjected to a stringent validation process using lysates from cells that are treated with pathway-specific activators and inhibitors to further confirm target specificity.

CONFIRMED LOT-TO-LOT CONSISTENCY

All Thunder™ kits are developed, validated and manufactured to ensure lot-to-lot consistency using a functional quality control assay where each new lot is compared to the previous lot.

AFFORDABILITY

Thunder™ is an affordable cell-based assay platform that provides easy access to the TR-FRET technology for all researchers looking to quantify low amounts of endogenous proteins in cells.

Assay workflow

All it takes is 3 simple steps to complete the workflow of all the TR-FRET Cell Signaling Assays.

Assays can be run using a 2-plate (transfer) protocol:

Or a 1-plate (all-in-one-well) protocol:

Assays are optimized to be run in 96-well or 384-well plates using the same total volume (20µL).

Typical validation data

Figure 2:Modulation of Phospho-IRβ (Y1150/Y1151) in B lymphocytes

Most popular targets

  • AKT (pan)
  • EGFR : Epidermal Growth Factor
  • STAT6 : Signal Transducer and Activator of Transcription 6
  • IL-2 & IL-6: Human Interleukin 2 & 6
  • IFNγ : Human Interferon Gamma
  • cAMP : adenosine 3′,5′-cyclic monophosphate

Discover all THUNDER kits available

  • Cell Signalling Assay Kits : Simple and rapid detection of either a specific phosphorylated target protein (Phospho-protein kit) and/or total protein in cell lysates (Total protein kit)
  • Biomarker Assay Kits : Assay developed to easy and quickly quantitate pro-inflammatory human cytokines in cell supernatants
  • cAMP Assay Kits : Assay kit provide a simple, rapide, robust and affordable tool to measure intracellular cAMP changes following G-protein coupled receptor (GPCR) activation.

Try a real alternative to perform your cell signalling studies in a different way!

Contact us to discuss your project
Cell Biology and Signalling, Supplying Discovery Tools

Cell signalling and STING studies – First cyclic GAMP immunoassay now available

26/07/2018 by Isabelle Nobiron, PhD No Comments

The innate immune system is the body’s crucial first defense against viral infections and it is dependent upon a group of pattern recognition receptors. cGMP-AMP Synthase (cGAS) responds to the presence of DNA in the cytosol by producing 2’,3’cGAMP. Cyclic GAMP in turn binds to STING leading to TBK1-mediated IRF3 activation and the production of type 1 IFN.

Understanding the STING cell signalling pathway and the role cGAMP plays in a number of important areas, such as infection, cancer, inflammation & senescence, relies on cGAMP measurement. Good news – Arbor Assays have just released a new cGAMP EIA kit, read on to learn more!

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Supplying Discovery Tools

Cell signalling – Discover G-LISA to study Small GTPases

21/06/2018 by Frédéric Samazan No Comments

As is already known, multiple families of small GTPases (Ras, Arf and Rho families) make up the Ras cell signalling - GTPase molecular cycleGTPase superfamily. Due to their role as molecular switches in cell signalling (active GTP-bound state vs inactive GDP-bound states) and in many other cellular responses (cytoskeletal reorganization, regulation of transcription, apoptosis…), these small GTPases are the subject of intense investigation to try to understand the mechanisms that regulate activation and inactivation of this proteins.

In this post, I invite you to discover a revolutionary way to study these cell signalling events by quantifying the level of active small GTPases in your experimental model.

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Cell Biology and Signalling

Cell signalling studies: Tips for selecting the best Small G Protein activation assay

25/04/2018 by Frédéric Samazan No Comments

Small GTP-binding proteins such as Rho and Ras GTPase family members are involved in regulating cell signalling pathways and impact a wide range of cellular processes, functions, and morphology. In this post, you’ll discover two types of Small G-protein Activation Assays to easily measure the GTP-bound form of your protein of interest, as well as some tips for choosing between both solutions and finding the assays that fit best with your cell signalling experiments.

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