VHL/CUL2/ELOB/ELOC/RBX1 complex of human VHL (GenBank Accession No. NM_000551) a.a. 2-212(end) with N-terminal FLAG-tag and MW= 25 kDa, human CUL2 (GenBank Accession No. NM_003591), a.a. 2-745(end) with N-terminal FLAG-tag and MW= 88 kDa, human ELOB (GenBank Accession No. NM_007108), a.a. 2-118 with N-terminal FLAG-tag and MW= 14 kDa, ELOC (GenBank Accession No. NM_005648), a.a. 2-112(end) with N-terminal His-tag and MW= 13 kDa human Rbx1 (GenBank Accession No. NM_014248), a.a. 2-108(end) with N-terminal His-tag and MW= 13 kDa, individually expressed in a HEK293 expression system.

VHL: P40337; CUL2: Q13617; ELOB: Q15370; ELOC: Q15369; Rbx1: P62877

VHL, CUL2, ELOB, ELOC, RBX1

Aqueous buffer solution.

ARV-771-mediated binding of VHL BPS #100373 to BET bromodomain was assessed using A-screen technology. Various amounts of VHL were incubated in 10-ul reaction with ARV-771 and GSTBRD3( BD2) BPS #31033 at room temperature for one hour. After this, FLAG-acceptor beads and GSH-donor beads were added followed by detection of A-counts.

40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 20% glycerol, 3 mM DTT.

At least 6 months at –80°C.

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

1. Kamura, T., et al., Science. 1999; 284(5414): 657-661.
2. Stebbins, C.E., et al., Science. 1999; 284(5413): 455-461.

Avoid freeze/thaw cycles.

Ubiquitin

Protein