AP2 | Floral homeotic protein APETALA 2

1 µg of total protein from Oryza sativa lines grown under drought stress conditions and over expressing protein DREB (AP2 domain, 285 aa, 31.35 kDa) (lines 1-5) and purified 100 ng of DREB (line 6), were separated on 12.5 % SDS-PAGE and blotted 4h to Nitrocellulose membrane. Blots were blocked with blocking agent 5% BSA or 1X TBS, 0.1% Tween-20 with 5% w/v non-fat dry milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 6 000 for 1h at RT or overnight at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 x 5 min in 1XTBST at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:8000 in for 1h at RT or max. 2h with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions. Exposure time was preferred between 20 to 30 seconds or max. 1 min.

Courtesy of Dr. Sunita Yadav, Indian Institute of Technology Kharagpur, India

This antibody was used in western blots on Lepidium appelianum and Lepidium campestre AP2 Protein expressed in E. coli cells. Applied dilution was 1: 6000.

Contains 0.02 % sodium azide.