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The pharmacological validation of the interaction of a ligand with its protein target is referred to as target engagement. Target engagement studies are crucial in drug discovery as they can provide confirmation that the compound(s) under study are indeed binding to the target protein at the intended site in order to be both effective and safe. Traditionally, target engagement has been assessed on purified proteins in buffer or deduced from downstream pathway effects such as post-translational modifications (PTMs) or enzyme substrate changes. Both of these approaches have severe limitations such as lack of a physiologically relevant environment in the case of purified proteins, or the inability to confidently attribute PTMs to one particular protein target due to the existence of multiple pathways controlling modification.
Even target engagement methodologies that do take place within a cellular environment such fluorescence resonance energy transfer (FRET) or bioluminescence resonance energy transfer (BRET) technologies have limited physiological relevance, since they require modifications to the protein and cloning.
The Cellular Thermal Shift Assay (CETSA®) is based on the change of the thermal denaturation profile of the native target protein in a cellular context that occurs following the binding of a ligand (i.e. a drug candidate). CETSA® is performed by incubating cells with the test compound, followed by heating of the compound-treated cells, and then by quantifying the remaining soluble target protein. The thermally stabilized protein-ligand complex can then detected by Alpha or HTRF® assay technology. Both technologies are no-wash, highly sensitive with a broad dynamic range, and provide fast time-to-results. CETSA® is a patented method and a licence from Pelago Bioscience is required to use the technology.
CETSA® can be combined with antibody pair screening technologies such as Alpha or HTRF allowing the study of thousands of compounds in a High-Throughput fashion.
CETSA® combined with a proximity-based assay such as Alpha or HTRF:
Alpha / HTRF CETSA® is a real game changer in that it combines an intracellular, physiologically relevant assay with high-throughput potential and amenability to automation.
爱游戏平台注册登录 and Pelago Biosciences currently offer fully validated Alpha CETSA® kits covering 13 different protein targets, including EGFR, BCL-2, NFκB, and p38 MAPK.
We also recommend an additional 40 AlphaLISA® SureFire® kits for protein targets that have been pre-validated by Pelago as CETSAble®. This pre-validation ensures that the protein target displays physical properties that make it amenable to thermal shift studies as well as the applicability of the Alpha assay in the context of CETSA®.
CETSA® is a patented method and the use of these kits with CETSA® is permitted under licence from Pelago Bioscience.
A CETSAble® assay has undergone validation as to the amenability of the protein target to heat-based denaturation studies as well as the usability of the approach in common cell lines. It will require further optimization in the end-user’s hands for their particular application and experimental conditions.
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Measure binding of small molecule drug candidates to target protein in a cellular environment with Alpha CETSA® target engagement assays. For use in combination with Pelago Bioscience's CETSA (Cellular Thermal Shift Assay) technology, this kit provides a convenient no-wash assay for detection and quantitation of small molecule binding to Cereblon, and can be used in hit identification, hit confirmation, and lead optimization studies for increased confidence and better prediction early in the drug discovery process. Please note that a valid CETSA subscription from Pelago Biosciences is required for use.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
Measure binding of small molecule drug candidates to target protein in a cellular environment with Alpha SureFire® CETSA® target engagement assays. For use in combination with Pelago Bioscience's CETSA® (Cellular Thermal Shift Assay) technology, this kit provides a convenient no-wash assay for detection and quantitation of small molecule binding to STING, and can be used in hit identification, hit confirmation, and lead optimization studies for increased confidence and better prediction early in the drug discovery process. Please note that a valid CETSA® subscription from Pelago Biosciences is required for use.
The AlphaLISA® SureFire® Ultra™ total ribosomal protein S6 (RPS6) assay is a sandwich immunoassay for quantitative detection of Ribosomal S6 (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total RIPK1 assay is a sandwich immunoassay for quantitative detection of RIPK1 (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total Ret assay is a sandwich immunoassay for quantitative detection of Total (phosphorylated and non-phosphorylated) Ret in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total PLC G2 assay is a sandwich immunoassay for quantitative detection of PLC G2 (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total PLC G1 assay is a sandwich immunoassay for quantitative detection of Total (phosphorylated and non-phosphorylated) PLC G1in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total PI3 Kinase p85 assay is a sandwich immunoassay for quantitative detection of PI3 Kinase p85 (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total MLKL assay is a sandwich immunoassay for quantitative detection of MLKL (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total IKKα assay is a sandwich immunoassay for quantitative detection of IKKα (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
Measure binding of small molecule drug candidates to target protein in a cellular environment with Alpha SureFire® CETSA® target engagement assays. For use in combination with Pelago Bioscience's CETSA® (Cellular Thermal Shift Assay) technology, this kit provides a convenient no-wash assay for detection and quantitation of small molecule binding to CDK4, and can be used in hit identification, hit confirmation, and lead optimization studies for increased confidence and better prediction early in the drug discovery process. Please note that a valid CETSA® subscription from Pelago Biosciences is required for use.
Measure binding of small molecule drug candidates to target protein in a cellular environment with Alpha SureFire® CETSA® target engagement assays. For use in combination with Pelago Bioscience's CETSA® (Cellular Thermal Shift Assay) technology, this kit provides a convenient no-wash assay for detection and quantitation of small molecule binding to CDK2, and can be used in hit identification, hit confirmation, and lead optimization studies for increased confidence and better prediction early in the drug discovery process. Please note that a valid CETSA® subscription from Pelago Biosciences is required for use.
The AlphaLISA® SureFire® Ultra™ total KAP1 assay is a sandwich immunoassay for quantitative detection of KAP1 (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total FGF Receptor 2 assay is a sandwich immunoassay for quantitative detection of FGF Receptor 2 (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total FGF Receptor 1 assay is a sandwich immunoassay for quantitative detection of FGF Receptor 1 (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total STAT1 assay is a sandwich immunoassay for quantitative detection of STAT1 (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total JNK1/2 assay is a sandwich immunoassay for quantitative detection of JNK1/2 (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total eiF4E assay is a sandwich immunoassay for quantitative detection of eiF4E (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total c-myc assay is a sandwich immunoassay for quantitative detection of c-myc (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total TBK1 assay is a sandwich immunoassay for quantitative detection of TBK1 (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total Acetyl CoA carboxylase assay is a sandwich immunoassay for quantitative detection of Acetyl CoA carboxylase (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
The AlphaLISA® SureFire® Ultra™ total STAT6 assay is a sandwich immunoassay for quantitative detection of STAT6 (phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be a normalization phosphorylation studies.
This kit has been pre-validated for use in combination with Pelago Bioscience’s CETSA® (Cellular Thermal Shift Assay) technology. Please note that further assay optimization will be required in order to adapt the kit to your particular cell line and experimental conditions. A valid CETSA® licence from Pelago Bioscience or a single kit agreement is required for using the CETSA® technology.
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