Ub-Rhodamine 110

Ub-Rhodamine 110 is a quenched, fluorescent substrate for deubiquitinating enzymes. Cleavage of the amide bond between the C-terminal glycine of ubiquitin and rhodamine110 results in an increase in rhodamine fluorescence at 535 nm (Exc. 485 nm).

Additional Information

Product Name: Ub-Rhodamine 110
Also Known As: N/A
Catalog No.: M3021
Size: 5x50µg
Molecular Weight: 8936 Da by MS
Species: Human
Source: Synthetic
Stock: 20 mM Tris, 150 mM NaCl, 2 mM βME, 10% Glycerol
Concentration: N/A
Quality Assurance: ≥95% by RP-HPLC and SDS-PAGE
Storage: Powder at −20°C; solution at −80°C. Avoid multiple freeze/thaw cycles.
PDF Data Sheet: PDF DatasheetMSDS
NCBI RefSeq: N/A
Image(s): (Click image to enlarge)

Coomassie-stained SDS-PAGE
Lane 1: Molecular weight markers
Lane 2: Ub-Rhodamine 110
Activity assay of Ub-Rhodamine 110 (100 nM) with varying concentrations UCH-L3 (5 - 313 pM).
Reaction buffer: 20 mM Tris, pH 7.1 at 37°C, 150 mM NaCl, 2 mM DTT. Reactions contained 0.5 μM Ub-Rhodamine 110 alone (control, brown line), or with 10 nM GST-Usp15 (gray line) or with 20 nM GST-Usp15 (yellow line). Rhodamine110 fluorescence was monitored by using a plate reader with excitation/emission filters at 485/20 and 530/30 nm, respectively.
Shipping Method: Dry ice shipping
References: 1. A. Tirat et al., (2005) Anal. Biochem., 343, 244-255.
2. U. Hassiepin et al., (2007) Anal. Biochem., 371, 201-207.

Details

Ub-Rhodamine 110 is a quenched, fluorescent substrate for deubiquitinating enzymes. Cleavage of the amide bond between the C-terminal glycine of ubiquitin and rhodamine110 results in an increase in rhodamine fluorescence at 535 nm (Exc. 485 nm).

Sample Preparation:
Dilute stock solution into required buffer. Typical reaction concentrations vary from 0.1 - 1 µM

Images:
(Click image to enlarge)

Coomassie-stained SDS-PAGE
Lane 1: Molecular weight markers
Lane 2: Ub-Rhodamine 110
Activity assay of Ub-Rhodamine 110 (100 nM) with varying concentrations UCH-L3 (5 - 313 pM).
Reaction buffer: 20 mM Tris, pH 7.1 at 37°C, 150 mM NaCl, 2 mM DTT. Reactions contained 0.5 μM Ub-Rhodamine 110 alone (control, brown line), or with 10 nM GST-Usp15 (gray line) or with 20 nM GST-Usp15 (yellow line). Rhodamine110 fluorescence was monitored by using a plate reader with excitation/emission filters at 485/20 and 530/30 nm, respectively.