Anti-AGO1e | Argonaute 1e (Zea mays)

Samples:
M– Marker Wild type (WT)
1 – 35 ug  of Zea mays premeiotic anthers extract
2 ­ - 35 ug of Zea mays unpollinated ovaries extract ago1e mutant
3 – 36 ug of Zea mays premeiotic anthers extract
4 -  ­ 35 ug of Zea mays unpollinated ovaries extract
M – Marker

Total protein extracted from Zea mays premeotic anthers. Samples were separated at RT using Tris-Acetate 3-8% NuPage LDS-PAGE gels, then blotted to PVDF membrane (0.45um pore size) using wet transfer O/N in the cold. Blot was stained with Ponceau and dried down. Blot was blocked with 5% milk in TBS-T for 1 hour at RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:500 with 2% milk in 5mL TBS-T O/N in the cold with agitation. Blot was washed with TBS-T 3x at RT with agitation (5 min per wash). Blot was incubated in matching secondary antibody (Goat anti-rabbit IgG (H&L), HRP conjugated, AS09 602, Agrisera) diluted to 1:25 000 in 2% milk in TBS-T for 1 hr at RT with agitation. The blot was washed with TBS-T 3x at RT with agitation (5 min per wash), then developed with the Agrisera ECL Bright and SuperBright reagents. Exposure time was 30 seconds.

Courtesy Dr. María Ximena Anleu Gil, Meyers Lab | UC Davis Genome Center, USA