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ChoosE2-Opti™ Intrachain TR-FRET Assay Kit
ChoosE2-Opti™ Intrachain TR-FRET Assay Kit
Artikelnummer
BPS78561
Verpackungseinheit
384 rxns.
Hersteller
BPS Bioscience
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Applications: Identify E2 enzymes that are partners of an E3 ligase of interestAssess the activity of new E2-E3 protein pairsScreen new E3 ligases and variants in HTS applicationsScreen molecules that inhibit/activate E2 enzyme activity in HTS applications,Determine compound IC50Analyze E3 ligase activity in real-time
Background: Covalent conjugation to ubiquitin (Ub) is one of the major post-translational modifications that regulates protein stability, function, and localization. Ubiquitination is the concerted action of three enzymes: a Ub-activating enzyme (E1), a Ub-conjugating enzyme (E2), and a Ub ligase (E3). The specificity and efficiency of ubiquitination are largely determined by the E2-E3 complex where the E3 enzyme directs the last step of the Ub-conjugating cascade by binding to both an E2~Ub conjugate and a substrate protein.
Contraindications: ChoosE2-Opti™ Intrachain TR-FRET Assay Kit is not suitable for measuring mono-ubiquitination. Weak signals may be obtained for multi-mono-ubiquitination or for short poly-ubiquitin chains.This kit is compatible with up to 1% final DMSO concentration. We recommend preparing the inhibitor in no higher than 5% DMSO solution in assay buffer and using 4 µl per well.The E3 ligase used in the assay should be purified. The assay will not perform well when using cell lysates due to the presence of other E3 ligases and ubiquitin.
Description: ChoosE2-Opti™ Intrachain TR-FRET Assay Kit is a sensitive high-throughput screening (HTS) TR-FRET Assay Kit, designed to identify E2 enzyme(s) that are functional partners for a purified E3 ubiquitin ligase of interest and lead to its self-polyubiquitination. The assay kit contains five E2 enzymes to be tested with the E3 ligase of interest, and two E3 ligases (SMURF1 and MDM2) to use as internal controls. It also contains an E1 enzyme, ATP, an optimized TRF Ubiquitin Mix, and a universal buffer. It utilizes a Europium-labeled Ubiquitin Donor as well as Cy5-labeled Ubiquitin Acceptor to complete the TR-FRET pairing. Since both the TR-FRET donor and acceptor are incorporated into polyubiquitin chains, this FRET-based assay requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time kinetics analyses.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Supplied As: ChoosE2-Opti™ Intrachain TR-FRET Assay Kit comes in a convenient 384-well reaction format.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
Applications: Identify E2 enzymes that are partners of an E3 ligase of interestAssess the activity of new E2-E3 protein pairsScreen new E3 ligases and variants in HTS applicationsScreen molecules that inhibit/activate E2 enzyme activity in HTS applications,Determine compound IC50Analyze E3 ligase activity in real-time
Background: Covalent conjugation to ubiquitin (Ub) is one of the major post-translational modifications that regulates protein stability, function, and localization. Ubiquitination is the concerted action of three enzymes: a Ub-activating enzyme (E1), a Ub-conjugating enzyme (E2), and a Ub ligase (E3). The specificity and efficiency of ubiquitination are largely determined by the E2-E3 complex where the E3 enzyme directs the last step of the Ub-conjugating cascade by binding to both an E2~Ub conjugate and a substrate protein.
Contraindications: ChoosE2-Opti™ Intrachain TR-FRET Assay Kit is not suitable for measuring mono-ubiquitination. Weak signals may be obtained for multi-mono-ubiquitination or for short poly-ubiquitin chains.This kit is compatible with up to 1% final DMSO concentration. We recommend preparing the inhibitor in no higher than 5% DMSO solution in assay buffer and using 4 µl per well.The E3 ligase used in the assay should be purified. The assay will not perform well when using cell lysates due to the presence of other E3 ligases and ubiquitin.
Description: ChoosE2-Opti™ Intrachain TR-FRET Assay Kit is a sensitive high-throughput screening (HTS) TR-FRET Assay Kit, designed to identify E2 enzyme(s) that are functional partners for a purified E3 ubiquitin ligase of interest and lead to its self-polyubiquitination. The assay kit contains five E2 enzymes to be tested with the E3 ligase of interest, and two E3 ligases (SMURF1 and MDM2) to use as internal controls. It also contains an E1 enzyme, ATP, an optimized TRF Ubiquitin Mix, and a universal buffer. It utilizes a Europium-labeled Ubiquitin Donor as well as Cy5-labeled Ubiquitin Acceptor to complete the TR-FRET pairing. Since both the TR-FRET donor and acceptor are incorporated into polyubiquitin chains, this FRET-based assay requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time kinetics analyses.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Supplied As: ChoosE2-Opti™ Intrachain TR-FRET Assay Kit comes in a convenient 384-well reaction format.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
| Artikelnummer | BPS78561 |
|---|---|
| Hersteller | BPS Bioscience |
| Hersteller Artikelnummer | 78561 |
| Green Labware | Nein |
| Verpackungseinheit | 384 rxns. |
| Mengeneinheit | PAK |
| Produktinformation (PDF) | Download |
| MSDS (PDF) |
|
Immer für Sie da
Gernot Ensinger, M.Sc.
Laurent Haze
