Anti-IRT1 | Iron regulated transporter 1

Product no: AS11 1780
AS11 1780 | Clonality: Polyclonal | Host: Rabbit | Reactivity:Arabidopsis thaliana, Tagetes erecta
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  • Product Info
  • Immunogen:

    KLH-conjugated synthetic peptide derived from Arabidopsis thaliana IRT1 sequence, Q38856, At4g19690

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Immunogen affinity purified serum in PBS pH 7.4.
    Format: Lyophilized
    Quantity: 50 µg
    Reconstitution: For reconstitution add 50 µl of sterile water
    Storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles, This antibody can be stored in a solution containg 50 % glycerol, final concentration, Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    Tested applications: Western blot (WB)
    Recommended dilution: 1 : 5000 (WB)
    Expected | apparent MW: 36.7 kDa
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana
    Predicted reactivity: Brassica napus, Brassica oleracea, Camelina sativa, Capsella rubella, Noccaea caerulescens, Thlaspi cerulescens

    Species of your interest not listed? Contact us
    Not reactive in: Horderum vulagre, Solanum lycopersicum, Tagetes erecta
  • Application Examples
  • Application example

     

    western blot detection using anti-IRT1 antibody

    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

    western blot using anti-IRT1 antibodies

    60 μg of total protein from Arabidopsis thaliana wild type (Col- 0) and IRT1 over-expressor(OE.IRT1) extracted with Celytic buffer (C2360 Sigmaaldrich) were separated on 15 % SDS- PAGE and blotted 30 min. to PVDF turbo-blot membrane. 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 Over night at 4°C 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 5 min in TBST at RT with agitation. Blot was incubated in secondary antibody (AS09 602, anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera ) diluted to 1:10 000 in for 1h at RT with agitation. The blot was washed as above and developed for 15 min with chemiluminescent detection reagent: AgriseraECL Bright. Exposure time was 1 minute. Hoagland medium contained 50 μM Fe.

    Courtesty Haitham Ayeb, Louvain Institute of Biomolecular Science and Technology (LIBST), Belgium
  • Background
  • Background:

    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.

  • Product Citations
  • Selected references: Cao et al. (2024). Spatial IMA1 regulation restricts root iron acquisition on MAMP perception. Nature. 2024 Jan;625(7996):750-759.  
    Domka et al. (2023) Endophytic yeast protect plants against metal toxicity by inhibiting plant metal uptake through an ethylene-dependent mechanism. Plant Cell Environ. 2023;46(1):268-287. doi:10.1111/pce.14473
    Kostic et al. (2022),The Relative Sensitivity of Marigold vs. Tomato to Iron (Fe) Toxicity Is Associated with Root Traits: Root-to-Shoot Mass Ratio, Failure to Sequester Fe in Roots, and Levels of the Major Fe-Uptake Protein, IRT, Horticulturae 2022, 8(9), 803;.
    Spielmann et al. (2022) Differential metal sensing and metal-dependent degradation of the broad spectrum root metal transporter IRT1. Plant J. 2022;112(5):1252-1265. doi:10.1111/tpj.16010.
    Gautam et al. (2021) IRONMAN Tunes Responses to Iron Deficiency in Concert with Environmental pH. bioRxiv 2021.02.16.431461; doi: https://doi.org/10.1101/2021.02.16.431461
    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.
    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.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

  • Reviews:
  • Reviews
    Below, you can grade the product on a scale from 0 to 5.
    Please also provide information about species, application, dilution and obtained result for the reviewed antibody.
    Your name will be displayed as the sender.
    Number of reviews: (2)
    Luisa M. Sandalio | 2021-07-13
    0 | 0
    This antibody works quite nicely, without any unspecificed bands in Arabidopsis roots
    Jeeyon Jeong | 2019-11-14
    0 | 0
    Species: Arabidopsis thaliana
    Application: Western blot
    Load per well: 20 g
    Dilution: 1:5000
    Result: Specific band at 35 kDa with no other non-specific signal when imaged with the Odyssey Licor system
    Affiliation: Amherst College

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AS16 ECL-S | extreme low femtogram detection  |  Limited stock

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