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Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: PROTAC ternary complex formation with BRD3 and cereblon was assessed using AlphaScreen™ technology using FLAG-acceptor beads and GSH-donor beads. 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.
Description: 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.
Format: Aqueous buffer solution
Formulation: 40 mm Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 0.04% Tween-20 and 20% Glycerol.
Genbank: Cereblon: NM_016302; DDB1: NM_001923; Cul4a: NM_003589; Rbx1: NM_014248
Storage Stability: At least 6 months at -80°C.
Supplied As: Aqueous buffer solution.
Tags: Cereblon: N-terminal FLAG-tag; DDB1: N-terminal FLAG-tag; Cul4a: N-terminal His-tag; Rbx1: N-terminal His-tag
Uniprot: Cereblon:Q96SW2; DDB1: Q16531; Cul4a: Q13619; Rbx1: P62877
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Gang, Lu., et al., Science. 2014; 343(6168): 305-309.2. Zhu, Y.X., et al., Blood. 2011; 118: 4771-4779.
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