Human Casitas B-lineage lymphoma proto-oncogene b (MDM2) is an E3 ubiquitin-protein ligase that functions as a negative regulator of T-cell activation. It is a potential drug target in cancer immunotherapy. The MDM2 TR-FRET Assay Kit is designed to measure MDM2 auto-ubiquitination activity in a homogeneous 384 reaction format. It utilizes biotin-labeled ubiquitin and a terbium-labeled anti-GST antibody 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.

Mdm2,mouse double minute 2, human homolog, Hdm2

The homogeneous 384 reaction format

Catalog #	Component	Amount	Storage
80301	UBE1 (E1)	25 µg	-80°C	Avoid freeze/ thaw cycles!
80314	UBCH5b (E2)	200 µg	-80°C
80751	Human MDM2 (E3), GST-tag	10 µg	-80°C
	Biotin-Ubiquitin	450 µl	-80°C
	ATP (400 µM)	150 µl	-80°C
	MDM2 assay buffer	2x 5ml	-80°C
	Tb-labeled donor	10 µl	-20°C
	Dye-labeled acceptor	10 µl	-20°C
	Detection buffer	5 mL	-20°C
	White, nonbinding Corning, low volume microtiter plate	1	Room temp.

See assay protocol for detailed instructions.

At least 6 months from date of receipt when stored as directed.

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

1. Wienken, M. et al., J. Mol. Cell Biol. 2017; 9(1): 74-80.
2. Shaikh, M.F. et al., Ann. Clin. Lab. Sci. 2016; 46(6): 627-34.

Avoid freeze/thaw cycles.

Ubiquitin

Assay Kit