CBL-B TR-FRET Assay Kit

Catalog #

79575

$2,200 *

Size: 384 reactions

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

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 our Ubiquitination Screening & Profiling Services.

CBL-B TR-FRET

Figure: Illustration of Assay Principle

Need us to run inhibitor screens or profile your compounds against CBL-B? Check out our Ubiquitination Screening Services.

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Synonyms

UBE1, UBA1, ubcH5b, UBE2D2, E3 ubiquitin-protein ligase CBL-B, RING finger protein 56, CBL, CBLB, cbl TR-FRET

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Product Data Gallery

Product Info

Storage and Usage

Citations

Assay Kit Format

TR-FRET

Supplied As

The CBL-B TR-FRET Assay Kit is designed to measure CBL-B auto-ubiquitination activity in a homogeneous 384 reaction format.

Materials Required But Not Supplied

Fluorescent microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET)
Adjustable micropipettor and sterile tips

Format

Catalog #	Component	Amount	Storage
80301	UBE1 (E1)*	10 µg	-80 °C	Avoid freeze/ thaw cycles!
80314	UBCH5b (E2)*	100 µg	-80 °C
80415	Human CBL-B (E3), GST-tag*	10 µg	-80 °C
	Biotin-Ubiquitin	400 µl	-80 °C
	ATP (400 μM)	150 µl	-80 °C
	U2 assay buffer**	2 x 10 ml	-80 °C
	Tb-labeled donor	10 µl	-20°C
	Dye-labeled acceptor	10 µl	-20°C
79969	White 384-well microtiter plate	1	Room Temp.

*The concentrations of E1, E2, and E3 enzymes are lot-specific and will be indicated on the tubes containing the enzyme.

UniProt #

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

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.

References

1. Jeon, M.S., et al., Immunity. 2004; 21(2): 167-177

2. Chiang, Y.J. et al., Nature. 2000; 403: 216-220

Storage/Stability

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.

Applications

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

Shipping Temperature

-80°C

Warnings

Avoid freeze/thaw cycles.

Disclaimers and Limitations

BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.

End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.

Scientific Category

Ubiquitination

Regulatory Status

Research Use Only