IRT1 | Iron regulated transporter 1
AS11 1780 | Clonality: Polyclonal | Host: Rabbit | Reactivity:Arabidopsis thaliana
|Info:||More information||Add review|
|Recommended dilution||1 : 5000 (WB)|
|Expected | apparent MW||
|Confirmed reactivity||Arabidopsis thaliana|
|Predicted reactivity||Noccaea caerulescens, Thlaspi cerulescens|
|Not reactive in||Horderum vulagre|
|Selected references||Selote et al. (2014). Iron-binding E3 ligase mediates iron response in plants by targeting bHLH transcription factors. Plant Physiol. 2014 Dec 1. pii: pp.114.250837.
Ivanov et al. (2014). SORTING NEXIN1 Is Required for Modulating the Trafficking and Stability of the Arabidopsis IRON-REGULATED TRANSPORTER1. Plant Cell. 2014 Mar 4.
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
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