Modified
Dec 2012
RW

 

FFE_01

Advances in Free Flow Electrophorsis since 35 years

Top Stories, recent application developments

    a) Separation of Thylakoid membranes from Arabidopsis thaliana

      Part I: The importance of sample preparation

One part of the sample was prepared with 1% DDM (Dodecylmaltoside), the other part was solubilized with 1% Digitonin solution, followed by a centrifugation step at 10°C for ten Minutes.

Subsequently, each supernatant was applied to the FFE, using the interval Zone Electrophoresis (iZE) mode.

Thylakoi_scheme 3

The FFE separated sample was collected and Blue Native Gels (BN-PAGE) were run.

Thylakoi_scheme 4

Finally SDS-PAGE gels were also used for the visualization of the separations..

Thylakoi_scheme 6

Conclusion:
The sample preparation step is crucial for the successful separation of Thylakoid membranes. We observed big differences in the BN and SDS pattern in the comparison of DDM and Digitonin based solubilization. The proteins will be further analyzed by mass spectrometry and published shortly.

Publication pending (Eichacker, et al. 2013)