His ub variants

Manufactured by R&D Systems

His-Ub variants are recombinant proteins that contain a polyhistidine tag (His-tag) fused to the N-terminus of ubiquitin. These proteins are used as tools in biochemical and cell-based research applications.

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Lab products found in correlation

E2 ubch5a, by R&D Systems (1 mentions) Nedd4, by Merck Group (1 mentions) E1 ube1, by R&D Systems (1 mentions) E2 ubch5a e2 616, by R&D Systems (1 mentions)

2 protocols using his ub variants

In Vitro Ubiquitination of PTEN

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For in vitro ubiquitination, 400 ng of purified Flag-PTEN from 293 cells was incubated with 40 ng E1 (UBE1) (E-305 Boston Biochem), 500 ng E2 (UbcH5a) (E2–616 Boston Biochem), 10 μg His-Ub variants (Boston Biochem), 8 mM ATP, 1× ligase reaction buffer [Fisher Scientific (Thermo Fisher Scientific) # B71], and 1× Energy regeneration system (Boston Biochem #B-10), 400 ng NEDD4 (Sigma Aldrich # SRP0226), and 400 ng WWP1 (Sigma Aldrich # SRP0229) at 37°C for 1 hour in 25 μl of reaction mixture.

Lee Y.R., Chen M., Lee J.D., Zhang J., Lin S.Y., Fu T.M., Chen H., Ishikawa T., Chiang S.Y., Katon J., Zhang Y., Shulga Y.V., Bester A.C., Fung J., Monteleone E., Wan L., Shen C., Hsu C.H., Papa A., Clohessy J.G., Teruya-Feldstein J., Jain S., Wu H., Matesic L., Chen R.H., Wei W, & Pandolfi P.P. (2019). Reactivation of PTEN tumor suppressor for cancer treatment through inhibition of a MYC-WWP1 inhibitory pathway. Science (New York, N.Y.), 364(6441), eaau0159.

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In Vitro Nanog Ubiquitination Assay

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For in vitro Nanog ubiquitination, 0.5 mg of purified GST-Nanog WT or ΔPDSST mutant from E. coli bacteria was incubated with 40 ng E1 (UBE1) (E-305 Boston Biochem), 0.5 mg E2 (UbcH5a) (E2–616 Boston Biochem), 10 mg His-Ub variants (Boston Biochem), 8 mM ATP, 1× ligase reaction buffer (Fisher Scientific (Thermo Fisher Scientific) # B71), and 1× Energy regeneration system (Boston Biochem #B-10), 0.5 mg E3 complex (Cullin 3/Rbx1/SPOP) at 37°C for 1 hour in 25 ml reaction mixture.

Zhang J., Chen M., Zhu Y., Dai X., Dang F., Ren J., Ren S., Shulga Y.V., Beca F., Gan W., Wu F., Lin Y.M., Zhou X., DeCaprio J.A., Beck A.H., Lu K.P., Huang J., Zhao C., Sun Y., Gao X., Pandolfi P.P, & Wei W. (2018). SPOP Promotes Nanog Destruction to Suppress Stem Cell Traits and Prostate Cancer Progression. Developmental cell, 48(3), 329-344.e5.

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