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AUX1 | Auxin transporter protein 1 (rabbit antibody)

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AS16 3159 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

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AS16 3159

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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.
Immunogen KLH-conjugated peptide derived from protein sequence of Arabidopsis thaliana AUX1. UniProt: Q96247, TAIR: AT2G38120
Host Rabbit
Clonality Polyclonal
Clone
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized
Quantity
Reconstitution
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 Collection of antibodies to auxins and proteins involved in auxin metabolism

AS11 1749| IAA | Auxin ELISA quantitation kit
AS16 3957 | Anti-AUX1 | Auxin transporter protein 1, goat antibody
Additional information
application information
Recommended dilution 1: 5000 (WB) on recombinant AUX1
Expected | apparent MW
Confirmed reactivity Arabidopsis thaliana (recombinant AUX1)
Predicted reactivity Arabidopsis thaliana
Not reactive in

no confirmed exceptions from predicted reactivity known in the moment

Additional information

Reactivity of this antibody on endogenous AUX1 remains to be determined.

Selected references

To be added when available, antibody released in December 2017.


Application example


Western blot using anti-AUX1 antibodies


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

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