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.
40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 20% glycerol, 3 mM DTT.
Species
Human
Application
Useful for the study of protein degradation and PROTAC design, and selectivity profiling.
Notes
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.