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IRT1 | Iron regulated transporter 1
- Product Info
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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
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Confirmed reactivity: Arabidopsis thaliana, Solanum lycopersicum, Tagetes erecta
Predicted reactivity: Brassica napus, Brassica oleracea, Camelina sativa, Capsella rubella, Noccaea caerulescens, Thlaspi cerulescens
Species of your interest not listed? Contact usNot reactive in: Horderum vulagre, Solanum lycopersicum, Tagetes sp. - Application Examples
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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
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
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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
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Selected references: 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
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Agrisera Western Blot protocol and video tutorials
Protocols to work with plant and algal protein extracts - Reviews:
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Luisa M. Sandalio | 2021-07-13This antibody works quite nicely, without any unspecificed bands in Arabidopsis rootsJeeyon Jeong | 2019-11-14Species: Arabidopsis thaliana
Application: Western blot
Load per well: 20 µg
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
Accessories
AS09 602 | Clonality: Polyclonal | Host: Goat | Reactivity: Rabbit IgG (H&L)
AS09 607 | Clonality: Polyclonal Host: Goat Reactivity: Rabbit IgG (H&L)
This product can be purchased in 3 different volumes:
AS16 ECL-S-10, 10 ml. Trial size limited to one per customer
AS16 ECL-S-100, 100 ml
AS16 ECL-S-1L, 1L
Choose the appropriate volume in the drop down menu to the right