File:Fpls-02-00018-g004.png

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English: Affinity purification of MP17:HIS. (A) Purification strategy of MP17:HIS; ATPMH2 rosettes were homogenized in NP40-buffer and soluble (1) and insoluble fractions were separated by centrifugation; the insoluble fraction was washed four times to reduce unspecific binding (last supernatant = 2); the insoluble fraction was subsequently extracted with urea and the urea insoluble (3) and soluble fraction (4) were separated by centrifugation; the soluble fraction was used for affinity purification on Nickel-NTA resin; fraction 5 represents the flow through and fraction 6 the last wash; bound proteins were eluted with EDTA (7); (B) Aliquots of the indicated purification steps (1–7) were separated by SDS-PAGE and stained with colloidal coomassie; (C) MP17:HIS (arrows) was detected by immuno blotting using anti-MP17-antiserum; WT = wild type protein extract.
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Source doi:10.3389/fpls.2011.00018
Author Katrin Link, Florian Vogel, Uwe Sonnewald

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current08:36, 6 June 2017Thumbnail for version as of 08:36, 6 June 20171,890 × 1,378 (2.91 MB)TIB-NOA (talk | contribs)pattypan 17.05

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