NRT1,1 | Nitrate transporter 1,1

AS12 2611 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

20 % until end of March 2023. Use discount code: Nitro20

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana NRT1.1 C-terminus, UniProt: Q05085, TAIR: AT1G12110

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 Immunolocalization (IL), Western blot (WB)

Recommended dilution 1 : 5000 (WB)

Expected | apparent MW

64 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Predicted reactivity Brassica napus
Species of your interest not listed? Contact us

Not reactive in Solanum tuberosum

Application examples

Application examples application example

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

Additional information

Background

Background NRT1.1 (nitrate transporter 1.1) is a dual-affinity nitrate transporter which acts also as a nitrate sensor. High-affinity nitrate transporter when phosphorylated and low-affinity transported when dephosphorylated. Low nitrogen concentration stimulates phosphorylation. Expressed in lateral roots.

Alternative names: AtNRT1, Nitrate/chlorate transporter, protein CHLORINA 1, AtNPF6.3.

Product citations

Selected references Medici et al. (2015). AtNIGT1/HRS1 integrates nitrate and phosphate signals at the Arabidopsis root tip. Nat Commun. 2015 Feb 27;6:6274. doi: 10.1038/ncomms7274

background:	NRT1.1 (nitrate transporter 1.1) is a dual-affinity nitrate transporter which acts also as a nitrate sensor. High-affinity nitrate transporter when phosphorylated and low-affinity transported when dephosphorylated. Low nitrogen concentration stimulates phosphorylation. Expressed in lateral roots. Alternative names: AtNRT1, Nitrate/chlorate transporter, protein CHLORINA 1, AtNPF6.3.
Picture (footer):	application example 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
All references:	Medici et al. (2015). AtNIGT1/HRS1 integrates nitrate and phosphate signals at the Arabidopsis root tip. Nat Commun. 2015 Feb 27;6:6274. doi: 10.1038/ncomms7274
Confirmed reactivity:	Arabidopsis thaliana
predicted reactivity:	Brassica napus Species of your interest not listed? Contact us
not reactive in:	Solanum tuberosum
calculated \| apparent molecular mass [kDa]:	64 kDa
Clonality:	Polyclonal
Format:	Lyophilized
Host:	Rabbit
immunogen:	KLH-conjugated synthetic peptide derived from Arabidopsis thaliana NRT1.1 C-terminus, UniProt: Q05085, TAIR: AT1G12110
Purity:	Immunogen affinity purified serum in PBS pH 7.4.
Quantity:	50 µg
recommended dilution:	1 : 5000 (WB)
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:	Immunolocalization (IL), Western blot (WB)