100X RELAY^TR Europium/Cy5 Ubiquitin Mix

Time resolved (TR)-FRET offers a sensitive, low background, stable and homogenous assay for determination of protein ubiquitination. Europium-ubiquitin and Cy5-ubiquitin are a pair of modified ubiquitin suitable for TR-FRET assays detecting formation of polyubiquitin chains and multi-monoubiquitination, thereby reporting the activity of E2 and E3 enzymes. The concentration and molar ratio of europium-ubiquitin and Cy5-ubiquitin in the RELAY^TR Europium/Cy5 Ubiquitin Mix are optimized to achieve excellent signal-to-blank ratio in E3 autoubiquitination assays. Typical protein concentration ranges are: UbE1 (10-30 nM), E2 (25-250 nM) and E3 (10-250 nM).

Reaction time is usually 1-3 hours in kinetic or end point assays.

Additional Information

Product Name:

Also Known As:

Eu-Ub; Eu-Ubiquitin; Cy5-Ub, Cy5-Ubiquitin

Catalog No.:

T3501-2K

Size:

2,000 x 20 µl Reactions

Molecular Weight:

Eu-Ub (9.5 kDa); Cy5-Ub (9.1 kDa)

Species:

Human

Source:

Bacterial recombinant

Stock:

Concentration:

See tube label

Quality Assurance:

~ 95% by SDS-PAGE; Validated by RELAY^TR ubiquitination assays.

Storage:

Store at -80°C; avoid multiple freeze-thaw cycles

PDF Data Sheet:

PDF Datasheet

NCBI RefSeq:

Image(s):

Images:
(Click image to enlarge)

A. Coomassie stained SDS-PAGE with 5 µg Terbium-Ub or Europium-Ub.
B. In-gel fluorescence visualization of Cy5-Ub in an SDS-PAGE.
C. Autoubiquitination of 25 nM CHIP detected by a RELAY^TR FRET assay with Eu-Ub and Cy5-Ub. The ratios (R) of fluorescence intensities at 655 nm and 620 nm from reactions containing E1/E2/Ub Mix/ATP with and without E3 are Signal (R_S) and Blank (R_B), respectively. The signal-to-blank ratio is calculated by using the formula of (R_S-R_B)/R_B. Time 0 is ~5 min after ATP addition. Data are mean±S.D. of triplicates.
D. Autoubiquitination of 12.5 nM MURF2 detected by a RELAY^TR FRET assay.
E. Autoubiquitination of 50 nM MDM2 detected by a RELAY^TR FRET assay.

Shipping Method:

Dry ice shipping

References:

1. https://www.bmglabtech.com/en/tr-fret/

Details

Time resolved (TR)-FRET offers a sensitive, low background, stable and homogenous assay for determination of protein ubiquitination. Europium-ubiquitin and Cy5-ubiquitin are a pair of modified ubiquitin suitable for TR-FRET assays detecting formation of polyubiquitin chains and multi-monoubiquitination, thereby reporting the activity of E2 and E3 enzymes. The concentration and molar ratio of europium-ubiquitin and Cy5-ubiquitin in the RELAY^TR Europium/Cy5 Ubiquitin Mix are optimized to achieve excellent signal-to-blank ratio in E3 autoubiquitination assays. Typical protein concentration ranges are: UbE1 (10-30 nM), E2 (25-250 nM) and E3 (10-250 nM).
Reaction time is usually 1-3 hours in kinetic or end point assays.

100X RELAYTR Europium/Cy5 Ubiquitin Mix

Additional Information

Details

100X RELAY^TR Europium/Cy5 Ubiquitin Mix