Products from BPS Bioscience require a minimum order value above 400€
Applications: Screen molecules that inhibit UbcH7 activity in drug discovery High-Throughput Screening (HTS) applications.
Background: UbcH7 (Ubiquitin-conjugating enzyme E2 L3 or UBE2L3) is an E2 ubiquitin-conjugating protein that receives Ubiquitin from a Ubiquitin-activating (E1) enzyme and subsequently interacts with a Ubiquitin ligase (E3) to conjugate Ubiquitin to substrate proteins and mediate their selective degradation. UbcH7 is involved in the ubiquitination of the transcription factors NF-κB, p53, and c-Fos, and is involved in cell cycle regulation. Abnormalities in UbcH7 activity has been linked to autoimmune diseases such as rheumatoid arthritis, celiac disease, Crohn's disease, and systemic lupus erythematosus. UbcH7 is, therefore, an attractive therapeutic target.
Contraindications: The UbcH6 TR-FRET Assay Kit is compatible with up to 1% final DMSO concentration. We recommend preparing the inhibitor in no higher than 10% DMSO in U2 Assay Buffer and using 2 µl per well.
Description: The UbcH7 (Ubiquitin-conjugating enzyme E2 L3 or UBE2L3) TR-FRET Assay Kit is a homogeneous, sensitive TR-FRET assay designed to measure UbcH7 ubiquitination activity in a 384-reaction format. It utilizes biotin-labeled Ubiquitin and a Terbium-labeled antibody recognizing the His-tagged UbcH7 protein to complete the TR-FRET pairing. This FRET-based assay requires no time-consuming washing steps, making it especially suitable for high throughput screening applications. The kit contains enough purified UbcH7 (full-length, His-Tag), purified UBE1, Biotin-Ubiquitin, anti-His Tb-labeled donor, dye-labeled streptavidin acceptor, and assay buffer for 400 reactions. The Terbium-labeled anti-His antibody binds to the His-tagged E2 conjugating protein, while the Dye-labeled streptavidin acceptor binds to Biotinylated-Ubiquitin. The complex forms when ubiquitin is transferred to the E2 enzyme, and the TR-FRET signal can be measured using a fluorescence plate reader capable of measuring Time Resolved-Fluorescence Resonance Energy Transfer.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
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