Anti-SQE1-3 | Squalene epoxidase1-3

20 µg/well of total protein freshly extracted from Withania somnifera and Arabidopsis thaliana have been used. Exact buffer components were: TrisHCl 100 mM, MgCl₂ 10 mM, NaEDTA 4 mM, 10% of glycerol and 1% of protease inhibitor solution; and denatured with exact buffer components at 95°C/5 min.  Samples were separated on 10 % SDS-PAGE and blotted for 7 minutes to PVDF (pore size of 0.2 µm), using a Trans-BlotTurbo Transfer System. The blot was blocked with 5% milk for/RT with agitation. The blot was incubated in the primary antibody at a dilution of 1: 500 in TBS-T ON/4°C with agitation. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS at RT with agitation. Blot was incubated in a matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 25 000 in TBS for 1 h/RT with agitation. The blot was washed as above and developed with the following chemiluminescent detection reagent: Agrisera ECL Bright. Exposure time was 45 seconds.
For lower background signal, primary antibody should be incubated 1h/RT at the dilution of 1: 1000.

Courtesy of Dr. Simone Landi, Università degli Studi di Napoli Federico II, Italy

Aletrnative names:Protein DROUGHT HYPERSENSITIVE 2, Squalene monooxygenase, XF1 protein