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. 2016 Apr 1;5(4):659–669. doi: 10.1002/mbo3.360

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© 2016 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Identification of Cry1Ac‐binding proteins on the BBMV from Helicoverpa armigera using the pull‐down method without elution. Proteins were resolved in PAGE discontinuous acrylamide gradients from 8% to 12%. Lanes 4 and 7 were divided into seven fractions (F‐1 to F‐7 and R‐1 to R‐7) and each fraction was used for LC‐MS/MS analysis. M: marker; 1: proteins from H. armigera BBMV incubated with CNBr‐Cry1Ac‐coupled agarose; 2: unbound proteins after the first incubation with CNBr‐Cry1Ac agarose; 5: unbound proteins after reincubation with CNBr‐Cry1Ac agarose; 3 and 6: proteins found in the elution obtained after last wash with phosphate‐buffered saline (PBS); 4 and 7: proteins bound to CNBr‐Cry1Ac after first and reincubations, respectively.