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Anti-NRT2,1 | Nitrate transporter 2,1
AS12 2612 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
- Product Info
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Immunogen: KLH-conjugated synthetic peptide derived from Arabidopsis thaliana NRT2.1 C-terminal sequence, UniProt: O82811, TAIR:AT1G08090
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. 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: 57.7 | 45 kDa - Reactivity
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Confirmed reactivity: Arabidopsis thaliana Predicted reactivity: Brassica juncea, Brassica napus, Brassuca rapa, Ricinus communis
Species of your interest not listed? Contact usNot reactive in: Alexandrium pacificum, Glycine max, Solanum tuberosum - Application Examples
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application information
50 µg of microsomal protein from wild type Arabidopsis thaliana (Col 0) roots at various stages of development (21 and 42 days) and from NRT2.1 deletion mutant (KO-NRT2.1) were solubilized with Laemmli x2 sample buffer and were separated on 12 % SDS-PAGE gel. The proteins were transferred (tank transfer, 100 V, 1h) to a PVDF membrane (Immobilon-P, 0.45 um, Millipore). Blot was blocked for 2h with agitation at room temperature in blocking buffer (SuperBlock, Thermo) containing 0.05 % Tween 20 (Thermo). Blot was then incubated for 2h at RT and with agitation in the fresh blocking buffer containing the primary antibody at a dilution of 1:5 000. The antibody solution was decanted and the blot was washed 3 times for 10 min in PBS-T (0.05 % Tween 20, Thermo) 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 blocking buffer, for 2h at RT with agitation. The blot was washed 1 time for 10 min in PBS-T and 2 times for 10 min in PBS. The blot was then incubated with ECL reagent (Super Signal West Pico,Thermo) for 5 min and the chemioluminescence was recorded on LAS 3000,Fuji. Exposure times were between 5-15 min using standard (lowest) sensitivity.
Courtesy of Dr. Wojciech Szponarski, French National Institute of Agricultural Research, France - Additional Information
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Additional information (application): Microsomal fraction is recommended to be used as it will have higher amount of NRT2,1 protein, If a total protein extract is used SDS should be applied from the beginning in extraction buffer as well as protease inhibitor coctail - Background
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Background: NRT2.1 is a high affinity nitrate transporter which is up-regulated by nitrate. Functions as a repressor of lateral root initiation independently of nitrate uptake. Synonymes: Protein ACH1, Protein LATERAL ROOT INITIATION 1, ACH1, ATNRT2.1, ATNRT2:1,, LIN1, NITRATE TRANSPORTER 2, NITRATE TRANSPORTER 2.1, NITRATE TRANSPORTER 2:1, NRT2, NRT2.1, NRT2:1, NRT2;1AT
- Product Citations
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Selected references: Zou et al. (2019). Phosphorylation at Ser28 stabilizes the Arabidopsis nitrate transporter NRT2.1 in response to nitrate limitation. J Integr Plant Biol. 2019 Jul 24. doi: 10.1111/jipb.12858. - Protocols
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Accessories
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