Molecular Weight Determination of DCB Proteins

MBP-tagged DCB^BIG1 proteins were resolved on a Superdex-75 HR10/300 analytical gel filtration column (GE Healthcare) at 0.5 ml/min in PBS (pH 7.5), 150 mM NaCl, 5 mM MgCl₂, and 0.001% (w/v) Triton X-100 before detection on a Wyatt Heleos II 18 angle light-scattering instrument coupled to a Wyatt Optilab rEX online refractive index detector and determination of native molecular weight (Perica et al., 2014 (link)).

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Galindo A., Soler N., McLaughlin S.H., Yu M., Williams R.L, & Munro S. (2016). Structural Insights into Arl1-Mediated Targeting of the Arf-GEF BIG1 to the trans-Golgi. Cell Reports, 16(3), 839-850.

Publication 2016

Superdex-75 HR10/300 Heleos II Optilab rEX

Gel filtration Light Mgcl2 Nacl Proteins Triton x 100

Corresponding Organization : MRC Laboratory of Molecular Biology

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Variable analysis

independent variables

The type of MBP-tagged DCB^BIG1 proteins used

dependent variables

Native molecular weight of the MBP-tagged DCB^BIG1 proteins

control variables

Flow rate of 0.5 ml/min
Buffer composition (PBS pH 7.5, 150 mM NaCl, 5 mM MgCl₂, 0.001% (w/v) Triton X-100)
Analytical gel filtration column (Superdex-75 HR10/300)
Detection methods (Wyatt Heleos II 18 angle light-scattering instrument coupled to a Wyatt Optilab rEX online refractive index detector)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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