Anti-SVR4 | Supressor of variegation 4

Application example


SVR4 antibody tested on thylakoids isolated from 1 week old barley. Thylakoids were isolated from 1 week old  Hordeum vulgare plants. The sample corresponding to 3ug Chl was separated on 12% Criterion XT Bis-Tris SDS-PAGE (BioRad) gels and blotted for 25min 100V to PVDF membrane. Blot was blocked with 5% fat free skimmed milk in PBS-T for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 overnight with agitation in 4˚C. The antibody solution was decanted and the blot was washed 3 times for 5 min in PBS-T at RT with agitation. Blot was incubated in secondary antibody (swine anti-rabbit IgG horse radish peroxidase conjugated, from Dako) diluted to 1: 5000 in 1% fat free skimmed milk in PBS-T for 1h at RT with agitation. The blot was washed 5 min in PBS-T and 1 min in PBS developed for 5 min with ECL according to the manufacturers instructions. Exposure time was 10 min.

Courtesy of Dr. Małgorzata Powikrowska, University of Copenhagen, Danmark

This protein is present in very low amounts only in early stages of plant development and this has to be taken into account when harvesting the tissue.

Western blots were done on: Arabidopsis thaliana total protein extract from cotyledons and Hordeum vulgare (2-14 days old plants).

SVR4 (Supressor of variegation 4) is a nuclear-encoded chloroplast-located protein required for proper function of the plastid transcriptional machinery. Synonymes: ATECB1, EARLY CHLOROPLAST BIOGENESIS 1, ECB1, MESOPHYLL-CELL RNAI LIBRARY LINE 7, MRL7.