Anti-IRT1 | Iron regulated transporter 1

Application example

5 µg of total protein from Arabidopsis thaliana wild type (Col-0) and IRT1 mutant (irt1-1) extracted with SDG buffer (62 mM Tris-HCL pH 8.6, 2.5 % SDS, 2 % dithiothreitol, 10 % glycerol) were separated on 15 % SDS-PAGE and blotted 1h to nitrocellulose. Blots were blocked with 5 % milk in TBST for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5 000 for 1h at RT with agitation in TBST with 2.5 % milk. The antibody solution was decanted and the blot was rinsed briefly three times, then washed once for 10 min in TBST at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:10 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturers instructions. Exposure time was 3 seconds.

Iron-sufficient medium contained 50 µM Fe, +Fe condition, iron-deficient medium 0 µM Fe, -Fe condition.

Courtesy Dr. Petra Bauer and Dr. Rumen Ivanov, Saarland University, Germany

IRT1 is a high afinity iron transported that plays a key role in the uptake of iron (in rhizosphere across the plasma membrane in the root epidermal layer) as well as mediates the heavy metals uptake under iron-defficiency. Is a principal regulator of iron homeaostasis in plants. Synonymes:Fe(2+) transport protein 1, Fe(II) transport protein 1.