Anti-LEA4-5 (1-77) | Late embryogenesis abundant protein LEA4-5 (N-terminal)

The amount of purified protein or peptide used in these dot western blot experiments is indicated above the image (2 ng to 200 ng). As specificity controls, we used 200 ng of purified AtLEA4-5 and of a peptide corresponding to the AtLEA4-5 C-region. In all cases, equal volumes of the different samples were dotted on Nitrocellulose Blotting Membrane 0.45 µm Amersham TM Protran TM Premium. Once samples were dry, membranes were blocked with non-fat milk 5% (W/V) during eight hours at 4°C with agitation. Blots were incubated with primary antibody at the indicated dilution for two days at 4°C with slow agitation. The antibody solution was decanted and blots were rinsed briefly, then washed once for 15 min and 2-times for 5 min in TBS-T at room temperature (RT) with agitation. Subsequently, blots were incubated with secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 for 8 h at RT with agitation. The blot was washed as described above and developed for 2 min with the chemiluminescent detection reagent. Exposure time was 60 seconds.

Courtesy of Dr. Alejandra A. Covarrubias,Universidad Nacional Autónoma de México, Mexico

LEA (Late embryogenesis abundant) proteins are very hydrophilic proteins, described over 25 years ago as accumulating during late stages of plant seed development. Found in vegetative plant tissues following exposure to environmental stress. Synonymes: Putative late embryogenesis abundant protein LEA.