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物种
Human分子别名
CUL4A/RBX1/DDB1/CRBN Complex表达序列
His-RBX1 (full-length), HA-DDB1 (full-length), Cullin4A (full-length), CRBN (full-length)表达宿主
Baculovirus-InsectCells纯度
＞90% by SDS-PAGE
内毒素含量
<1EU/μg标记
Unconjugated性状
Liquid缓冲体系
50 mM Tris, 200 mM NaCl, 2 mM TCEP, pH 8.0储存条件
Store at -80ºC. Avoid multiple freeze-thaw cycles.
Cullin-4A (CUL4A) is a core component of multiple cullin-RING type Ubiquitin ligase complexes that mediate the ubiquitination of proteins involved in cell cycle progression, DNA repair and other processes. In the DCX complex (DDB1-CUL4-X-box), CUL4A serves as a scaffold that organizes the DDB1-X-box substrate recognition subunits with the RBX1 subunit and contributes to catalysis through positioning of the substrate and an E2 ubiquitin-conjugating enzyme. In vivo, the E3 ubiquitin ligase activity of the DCX complex is dependent on neddylation of the CUL4 subunit, though neddylation may be dispensable in some in vitro reactions.
生物活性
The Cullin4A-RBX1-DDB1-CRBN activity was measured by protein-tracer binary binding assay using HTRF technology.
The Cullin4A-RBX1-DDB1-CRBN activity was measured by PROTAC-mediated Ternary Complex Formation using HTRF technology. The dose response curve of dBET1 in the Ternary Assay exhibited a bell-shaped curve, a hook effect and a peak.
The Cullin4A-RBX1-DDB1-CRBN activity was measured by WB analysis for target protein ubiquitination assay. The Ubiquitinated BRD4(D1+D2) bands increased with time.
电泳
Lane M: Precision Plus Protein™ Dual Color Standards (Bio-Rad) (Sample loaded: 5 μl)
Lane 1: Cullin4A-RBX1-DDB1-CRBN complex under reducing condition (Sample loaded: 2 μg)
Lane 2: Cullin4A-RBX1-DDB1-CRBN complex under reducing condition (Sample loaded: 4 μg)
体积排阻色谱(SEC-HPLC)
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