Cereblon/DDB1/Cul4A/Rbx1 complex of human Cereblon (GenBank Accession No. NM_016302) a.a. 2-442(end) with N-terminal FLAG-tag and MW= 51 kDa, human DDB1 (GenBank Accession No. NM_001923), a.a. 1-1140(end) with N-terminal FLAG-tag and MW= 128 kDa, human Cul4A (GenBank Accession No. NM_003589), a.a. 2-759(end) with N-terminal His-tag and MW= 87 kDa, human Rbx1 (GenBank Accession No. NM_014248), a.a. 2-108(end) with N-terminal His-tag and MW= 13 kDa, co-expressed in a HEK293 expression system.

Cereblon:Q96SW2; DDB1: Q16531; Cul4a: Q13619; Rbx1: P62877

CRBN, Protein X 0001, MRT2A MRT2

Aqueous buffer solution.

PROTAC ternary complex formation with BRD3 and cereblon was assessed using AlphaScreen™ technology. Various amounts of Cereblon were incubated in 10-µl reaction with 100 nM dBET1 and 15 nM GST-BRD3 BPS #31033 at room temperature for one hour. After this, FLAG-acceptor beads and GSHdonor beads were added followed by detection of A-counts.

40 mm Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 0.04% Tween-20 and 20% Glycerol.

At least 6 months at –80°C.

Useful for the study of protein degradation and PROTAC design, and selectivity profiling.

1.    Gang, Lu., et al., Science. 2014; 343(6168): 305-309.

2.    Zhu, Y.X., et al., Blood. 2011; 118: 4771-4779.

Avoid freeze/thaw cycles.

Ubiquitin

Protein