product information 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 Immunogen KLH-conjugated synthetic peptide derived from Arabidopsis thaliana NRT2.1 sequence, UniProt: O82811, TAIR:AT1G08090 Host Rabbit Clonality Polyclonal Clone Purity Affinity purified serum Format Lyophilized in PBS pH 7.4 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 tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes. Tested applications western blot (WB) Related products AS12 2611 | Anti-NRT1.1 | nitrate transporter 1.1, rabbit antibodyAS09 473| Anti-NRT1.4 | nitrate transporter, rabbit antibodycollection of antibodies for nitrogen metabolism Plant protein extraction buffer Secondary antibodies Additional information

application information
Recommended dilution		1 : 5000 with standard ECL (WB)
Expected | apparent MW		57.7 | 45 kDa
Confirmed reactivity		Arabidopsis thaliana
Predicted reactivity		Brassica juncea, Brassica napus, Brassuca rapa, Ricinus communis
Not reactive in		no confirmed exceptions from predicted reactivity are currently known
Additional information		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.
Selected references		to be added when available, antibody released in October 2014.

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

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