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CBL-B TR-FRET Assay Kit

Human Casitas B-lineage lymphoma proto-oncogene b (CBL-B) is an E3 ubiquitin-protein ligase that functions as a negative regulator of T-cell activation. It is a promising drug target in cancer immunotherapy. The CBL-B TR-FRET Assay Kit is designed to measure CBL-B auto-ubiquitination activity in a homogeneous 384 reaction format. It utilizes biotin-labeled Ubiquitin and a terbium-labeled antibody recognizing the GST-tagged CBL-B 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.Need us to run inhibitor screens or profile your compounds against CBL-B? Check out ourUbiquitination Screening & Profiling Services.Figure: Illustration of Assay PrincipleNeed us to run inhibitor screens or profile your compounds against CBL-B? Check out ourUbiquitination Screening Services.

Product Specifications

Background

CBL-B, a RING finger E3-ubiquitin ligase, acts as an intracellular checkpoint and a regulator of cytotoxic lymphocytes and NK cells. CBL-B binds E2-ubiqutin as well as target proteins such as TAM receptor TYRO3, forcing further degradation of TYRO3. CBL-B modulates innate immune responses and plays a key role in host defense to pathogens and anti-tumor immunity. Targeting CBL-B represents a potential therapeutic strategy for the management of human immune-related disorders such as autoimmune diseases, infections, tumors, and allergic airway inflammation. CBL-B has emerged as a new target of interest in the field of immunotherapy due to its ability to suppress cancer immune-evasion. CBL-B initiates anti-cancer immune responses in NK cells in both cytotoxic and effector T cells, promoting T-cell and NK cell activation.

UniProt

UBE1: P22314; UBCH5b: P62837; CBL-B: Q13191

Applications

Great for screening small molecular inhibitors for drug discovery and HTS applications.

Format

Catalog #ComponentAmountStorage80301UBE1 (E1) *10 µg-80 °CAvoid freeze/thaw cycles!80314UBCH5b (E2) *100 µg-80 °C80415Human CBL-B (E3), GST-tag*10 µg-80 °CBiotin-Ubiquitin400 µl-80 °CATP (400 μM) 150 µl-80 °CU2 assay buffer**2 x 10 ml-80 °CTb-labeled donor10 µl-20°CDye-labeled acceptor10 µl-20°C79969White 384-well microtiter plate1RoomTemp.*The concentrations of E1, E2, and E3 enzymes are lot-specific and will be indicated on the tubes containing the enzyme.

Shipping Conditions

-80°C

Storage Conditions

Stable at least 6 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
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