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His-RBX1 (full-length),
HA-Cullin2 (full-length),
HA-ElonginC (full-length),
ElonginB (full-length),
VHL (full-length)
＞90% by SDS-PAGE
\n\nLane M: Precision Plus Protein™ Dual Color Standards (Bio-Rad) (Sample loaded: 5 μl)
Lane 1: Cullin2-RBX1-ElonginB/C-VHL complex under reducing condition (Sample loaded: 2\nμg)
VHL (von Hippel-Lindau disease tumor suppressor) is the substrate recognition subunit for an E3 ligase activity that ubiquitinates proteins containing hydroxyproline residues. Targets of VHL include HIF1 alpha, beta 2 adrenergic receptor, ZHX2 and others. In VHL-box E3 Ubiquitin ligases, VHL is bound to a heterodimer complex composed of Elongin B (ELOB) and Elongin C (ELOC) through the "BC-box motif" found in many proteins in the VHL-box and SOCS-box protein families. ELOB/ELOC serves as an adapter between substrate recognition proteins and Cullin-2 (or Cullin-5) and RBX1 to form an active E3 ligase.
Store at -80ºC. Avoid multiple
freeze-thaw cycles.
参考图片
Lane M: Precision Plus Protein™ Dual Color Standards (Bio-Rad) (Sample loaded: 5 μl)
Lane 1: Cullin2-RBX1-ElonginB/C-VHL complex under reducing condition (Sample loaded: 2
μg)
The Cullin2-RBX1-ElonginB/C-VHL activity was measured by protein-tracer binary binding
assay using HTRF
technology.
The Cullin2-RBX1-ElonginB/C-VHL activity was measured by PROTAC-mediated Ternary
Complex Formation using HTRF
technology. The dose response curve of ARV-771 in the
Ternary Assay exhibited a bell-shaped curve, a hook effect and a peak.
The Cullin2-RBX1-ElonginB/C-VHL activity was measured by WB analysis for target protein
ubiquitination assay. The Ubiquitinated BRD4(D1+D2) bands increased with time.