U 550

Manufactured by R&D Systems

The U-550 is a programmable multi-channel pipette designed for precise and accurate liquid handling. It features an ergonomic design, adjustable volume settings, and digital display for easy operation. The device can accommodate a wide range of sample volumes, making it suitable for various laboratory applications.

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Lab products found in correlation

Ovalbumin (ova), by Merck Group (1 mentions) Infinite m1000 pro fluorometer, by Tecan (1 mentions) Envision plate reader, by PerkinElmer (1 mentions)

2 protocols using u 550

Ub-AMC Enzymatic Assays in Vitro

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In vitro enzymatic assays using ubiquitin-7-amido-4-methylcoumarin (Ub-AMC; U-550; Boston Biochem) were performed in 50–100 µl reaction buffer (20 mM HEPES-KOH [pH 7.8], 20 mM NaCl, 0.1 mg/ml ovalbumin [A7641; Sigma], 0.5 mM EDTA, and 10 mM DTT) at 25°C. Fluorescence was monitored in an Infinite® M1000 PRO Fluorometer (TECAN).

Deng M., Yang X., Qin B., Liu T., Zhang H., Guo W., Lee S.B., Kim J.J., Yuan J., Pei H., Wang L, & Lou Z. (2016). Deubiquitination and activation of AMPK by USP10. Molecular cell, 61(4), 614-624.

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Ubiquitin-AMC Hydrolysis Assay Protocol

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As described previously [30], Ub-AMC assays were conducted in a flat-bottom, low-flange 384-well plate in a 40-μl reaction with Ub AMC-conjugated proteins (Boston Biochem, U-550). Substrates and enzymes were incubated in Ub-AMC assay buffer (50 mM Tris-HCl, pH 7.5, 1 mM DTT, 1 mg/ml ovalbumin [RPI Corp, 9006-59-1], 1 mM EDTA, 5 mM MgCl₂, 1 mM ATP [New England BioLabs, P0756L]). After adding of Ub-AMC, the reaction was initiated, and an Envision plate reader (PerkinElmer, MA, USA) was used to measure mixture at Ex345/Em445. For determination of K_M, ZRANB1^T35D,S209D was mixed with the indicated concentrations of Ub-AMC. Lineweaver–Burk analysis was carried out using a linear regression fit of the data with the equation to calculate K_M: (1/ν) = K_M/V_max(1/[S]) + 1/V_max.

Feng X., Jia Y., Zhang Y., Ma F., Zhu Y., Hong X., Zhou Q., He R., Zhang H., Jin J., Piao D., Huang H., Li Q., Qiu X, & Zhang Z. (2019). Ubiquitination of UVRAG by SMURF1 promotes autophagosome maturation and inhibits hepatocellular carcinoma growth. Autophagy, 15(7), 1130-1149.

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