Protocol detail

Purification of Nedd8-related Proteins

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The following purified proteins were previously described in Kelsall et al., (2013 (link)): Nedd8 and the APPBP1–UBA3 heterodimer (Ohta et al., 2013 (link)); DAC–TEV–Cul3–RBX1 and KLHL3 (Schumacher et al., 2015 (link)); GST–CAND1. DCNL1, 2, 3, 4 and 5 were expressed with His₆ tags in BL21 and purified by Ni²⁺–Sepharose (GE Healthcare). GST–DCNL1 and GST–DCNL2 were purified by GSH–Sepharose (GE Healthcare).

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Keuss M.J., Thomas Y., Mcarthur R., Wood N.T., Knebel A, & Kurz T. (2016). Characterization of the mammalian family of DCN-type NEDD8 E3 ligases. Journal of Cell Science, 129(7), 1441-1454.

Publication 2016

Ni2+–Sepharose GSH–Sepharose

His6 tags Nedd8 Proteins Rbx1 Sepharose

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Variable analysis

independent variables

Nedd8
APPBP1–UBA3 heterodimer
DAC–TEV–Cul3–RBX1
KLHL3
GST–CAND1
DCNL1, 2, 3, 4 and 5 (with His6 tags)
GST–DCNL1
GST–DCNL2

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive control: None mentioned
Negative control: None mentioned

Annotations

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