Cul2/Rbx1 [untagged]

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Cul2/Rbx1 [untagged] is an insect cell-derived, functional human CUL2/RBX1 complex produced by coexpression in insect cells, validated indirectly through its ability to act as a substrate for neddylation in the presence of the NEDD8 E3 ligase His-DCNL2 and thioester-loaded His-UBE2M-NEDD8. Incubation of Cul2/Rbx1 and thioester loaded His-UBE2M- NEDD8 in the presence or absence of His-DCNL2 at 4°C was compared at two time points T0 and T2 minutes. Neddylation of the Cul2 subunit in the presence of His- DCNL2 was demonstrated.

Note

Cullin2 [Dac tagged]/Rbx1 was cleaved with TEV protease [6His tagged]. The Dac tag and TEV protease [6His-Tagged] were removed by capturing on amp sepharose and nickel resin respectively.

Specifications


• Species:                  human

• Source:                   insect (Sf21)

• Quantity:                25 μg

• Concentration:        0.5 mg/ml

• Formulation:          50 mM HEPES pH 7.5, 150 mM NaCl, 2 mM DTT, 10% glycerol

• Molecular weight:  Cul2 = ~87.1 kDa; Rbx1 = ~12.3 kDa

• Protein IDs:            confirmed by mass spectrometry

• Purity:                    >75% by InstantBlue^TM SDS-PAGE

• Stability/Storage:   12 months at -70°C/aliquot as required