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Fig. 9 | BMC Plant Biology

Fig. 9

From: Proanthocyanidin accumulation and transcriptional responses in the seed coat of cranberry beans (Phaseolus vulgaris L.) with different susceptibility to postharvest darkening

Fig. 9

Biochemical analysis of the recombinant PvANR1 enzyme. Coomassie Brilliant Blue stained SDS-PAGE (a) and immunoblot (b) analysis of the expression and purification of the recombinant PvANR1 from E. coli BL21 cells. With exception of lanes where corresponding protein amount is indicated, lanes were loaded by equal volume (15 μL) in a final of 1× SDS sample buffer. The immunoblot was subjected to a chromogenic stain following successive probing with an anti-His6 antibody and a secondary antibody conjugated to alkaline phosphatase. Abbreviations include: His6, hexahistidine; IMAC, immobilized metal affinity chromatography; IPTG, isopropyl β-D-thiogalactopyranoside; M r , relative molecular mass. c HPLC-DAD analysis of in vitro PvANR1 activity. The grey chromatogram represents authentic (−)-epicatechin and (+)-catechin standards. The blue chromatogram represents products formed from assays containing PvANR1, 100 μM cyanidin and 800 μM NADPH. The black chromatogram represents the analysis of an assay performed in the absence of PvANR1

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