AUX1 | Auxin transporter protein 1 (goat antibody)

AS16 3957 | Clonality: Polyclonal | Host: Goat | Reactivity: Arabidopsis thaliana

AUX1 | Auxin transporter protein 1 (goat antibody)

Product Information

Immunogen KLH-conjugated peptide derived from protein sequence of Arabidopsis thaliana AUX1. UniProt: Q96247, TAIR: AT2G38120

Host Goat

Clonality Polyclonal

Purity Immunogen affinity purified serum in PBS pH 7.4.

Format Lyophilized

Quantity 50 µg

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: 250 - 1: 700 (IL), 1: 5000 (WB) on recombinant AUX1

Reactivity

Confirmed reactivity Arabidopsis thaliana (recombinant AUX1)

Predicted reactivity Arabidopsis thaliana

Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Application example

Western blot using anti-AUX1 goat antibody

Arabidopsis thaliana AUX1 was expressed in insect cells using baculovirus infection. Expression was driven by the strong polyhedrin promoter. Cultures of Sf9 cells (20 ml, approx. 2 x 107 cells) were infected with virus at multiplicities of infection (MOI) of 0.1, 1 and 2.5. 2 mL samples were harvested by centrifugation after 3 days, lysed (20 mM Tris/HCl, pH 7.4, 200 mM NaCl, 1 mM EDTA, 1% Tween 20, protease inhibitors and DNAseI) at 4°C for 30 min, sonicated (3 x 5 s pulses) and centrifuged. Cleared supernatant samples (whole cell lysates) were run on SDS-PAGE using 8 – 13% acrylamide gradient gels. After transfer to PVDF, the membrane was blocked with TBS-Tween with 10% milk powder overnight. Primary antibodies were applied at 1:5000 dilution in TBS-Tween with 5% milk powder for one hour at room temperature, washed x3 in TBS-Tween for 10 mins each and secondary antibodies (e.g. AS09 605 rabbit anti-goat HRP conjugated, Agrisera) applied diluted 1:10 000 as above. After washing as above, development was by ECL using Millipore Immobilon Transfer reagent for 2 mins and the image captured by ImageQuant. As negative controls, samples of non-infected cells (NC) were run alongside the AUX1 extracts, as well as protein size markers (Mr).

The complete fusion protein (enhanced GFP-AtAUX1) was detected at close to 60 kDa A breakdown or truncated band was also seen at a little over 40 kDa.

Courtesty of Prof Richard Napier, University of Warwick, UK

Immunolocalization using anti-AUX1 antibodies

Whole mount tissue immunochemistry of sections of protophloems of Arabidopsis thaliana root tips.
Fixation: 4% PFA in MTSB Buffer pH 6.9 Vacum 2x5’ and 1x40’
Blocking: BSA in MTSB (0.4g in 20ml), As a secondary antibody Alexafluor 546 donkey, anti-goat, concentration 1:500.

Comparison between the two alleles: aux1-21 has a frame shift after exon 5, whereas aux1-2 only has an amino acid substitution and expression is checked in the protophloem.

Courtesy Ana Cecilia Aliaga Fandino ,Hardtke lab, University of Lausanne, Switzerland

Additional information

Reactivity of this antibody on endogenous AUX1 remains to be determined

Background

Background AUX1 (Auxin transporter protein 1) is a carrier protein involved in proton-driven auxin influx. Synthesized in developing leaves and transpported to tips. Involved in lateral root formation, trichoblast polarization and root hair elongation. Required for gravitropism and thigmotropism, especially in roots, by modulating responses to auxin, ethylene and cytokinins. Needed for ammonium-mediated root-growth inhibition. Alternative names: AUX1, AUXIN RESISTANT 1, WAV5, WAVY ROOTS 5, PIR1, MAP1, MODIFIER OF ARF7/NPH4 PHENOTYPES 1, Auxin influx carrier protein 1, Polar auxin transport inhibitor-resistant protein 1.

Product citations

background:	AUX1 (Auxin transporter protein 1) is a carrier protein involved in proton-driven auxin influx. Synthesized in developing leaves and transpported to tips. Involved in lateral root formation, trichoblast polarization and root hair elongation. Required for gravitropism and thigmotropism, especially in roots, by modulating responses to auxin, ethylene and cytokinins. Needed for ammonium-mediated root-growth inhibition. Alternative names: AUX1, AUXIN RESISTANT 1, WAV5, WAVY ROOTS 5, PIR1, MAP1, MODIFIER OF ARF7/NPH4 PHENOTYPES 1, Auxin influx carrier protein 1, Polar auxin transport inhibitor-resistant protein 1.
Clonality:	Polyclonal
Format:	Lyophilized
Host:	Goat
immunogen:	KLH-conjugated peptide derived from protein sequence of Arabidopsis thaliana AUX1. UniProt: Q96247, TAIR: AT2G38120
Purity:	Immunogen affinity purified serum in PBS pH 7.4.
Quantity:	50 µg
recommended dilution:	1: 250 - 1: 700 (IL), 1: 5000 (WB) on recombinant AUX1
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)
Confirmed reactivity:	Arabidopsis thaliana (recombinant AUX1)
predicted reactivity:	Arabidopsis thaliana
not reactive in:	No confirmed exceptions from predicted reactivity are currently known
additional information (application):	Reactivity of this antibody on endogenous AUX1 remains to be determined
Picture (footer):	Application example Arabidopsis thaliana AUX1 was expressed in insect cells using baculovirus infection. Expression was driven by the strong polyhedrin promoter. Cultures of Sf9 cells (20 ml, approx. 2 x 107 cells) were infected with virus at multiplicities of infection (MOI) of 0.1, 1 and 2.5. 2 mL samples were harvested by centrifugation after 3 days, lysed (20 mM Tris/HCl, pH 7.4, 200 mM NaCl, 1 mM EDTA, 1% Tween 20, protease inhibitors and DNAseI) at 4°C for 30 min, sonicated (3 x 5 s pulses) and centrifuged. Cleared supernatant samples (whole cell lysates) were run on SDS-PAGE using 8 – 13% acrylamide gradient gels. After transfer to PVDF, the membrane was blocked with TBS-Tween with 10% milk powder overnight. Primary antibodies were applied at 1:5000 dilution in TBS-Tween with 5% milk powder for one hour at room temperature, washed x3 in TBS-Tween for 10 mins each and secondary antibodies (e.g. AS09 605 rabbit anti-goat HRP conjugated, Agrisera) applied diluted 1:10 000 as above. After washing as above, development was by ECL using Millipore Immobilon Transfer reagent for 2 mins and the image captured by ImageQuant. As negative controls, samples of non-infected cells (NC) were run alongside the AUX1 extracts, as well as protein size markers (Mr). The complete fusion protein (enhanced GFP-AtAUX1) was detected at close to 60 kDa A breakdown or truncated band was also seen at a little over 40 kDa. Courtesty of Prof Richard Napier, University of Warwick, UK Whole mount tissue immunochemistry of sections of protophloems of Arabidopsis thaliana root tips. Fixation: 4% PFA in MTSB Buffer pH 6.9 Vacum 2x5’ and 1x40’ Blocking: BSA in MTSB (0.4g in 20ml), As a secondary antibody Alexafluor 546 donkey, anti-goat, concentration 1:500. Comparison between the two alleles: aux1-21 has a frame shift after exon 5, whereas aux1-2 only has an amino acid substitution and expression is checked in the protophloem. Courtesy Ana Cecilia Aliaga Fandino ,Hardtke lab, University of Lausanne, Switzerland