NRT1,1 | Nitrate transporter 1,1
AS12 2611 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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50 µg of microsomal protein from wild type Arabidopsis thaliana (Col 0) roots at various stages of development or from NRT1.1 deletion mutant (CHL1.5) were solubilized with Laemmli x2 sample buffer and were separated on 10 % SDS-PAGE gel. The proteins were transfered (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 of high sensitivity for 5 min and the chemioluminescence was recorded on LAS 3000,Fuji. Exposure times were between 1-10 min using standard (lowest) sensitivity.
Courtesy of Dr. Wojciech Szponarski, French National Institute of Agricultural Research, France
Alternative names: AtNRT1, Nitrate/chlorate transporter, protein CHLORINA 1, AtNPF6.3.
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