Journal: Plant Physiology
Article Title: Inositol 1,3,4,5,6-Pentakisphosphate 2-Kinase from Maize: Molecular and Biochemical Characterization [OA]
Figure Lengend Snippet: Detection of ZmIPK1 protein in different maize tissues via western blotting. Proteins were extracted from maize leaf (lane 1), immature ear (lane 2), root (lane 3), seed at 12 DAP (lane 4), endosperm at 15 (lane 5), 20 (lane 6), and 25 (lane 7) DAP, and embryo at 24 (lane 8) and 31 (lane 9) DAP. After SDS-PAGE on a 8% to 16% gradient Tris-Gly polyacrylamide gel, the proteins were transferred onto a nitrocellulose membrane, incubated with polyclonal antibodies against maize ZmIPK1 protein, and detected with goat anti-rabbit antibody conjugated to horseradish peroxidase. ZmIPK1 protein (10 ng) purified from E. coli lysate was used as the reference (lane 10).
Article Snippet: The unbound fractions containing ZmIPK1 protein were pooled and dialyzed against 20 m m Tris-HCl, pH 7.5, 1 m m DTT at room temperature for 2 h. Then, the sample was applied to an 8 mL Mono Q column (Amersham).
Techniques: Western Blot, SDS Page, Incubation, Purification