AUX1 | Auxin transporter protein 1 (rabbit antibody)
AS16 3159 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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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.
no confirmed exceptions from predicted reactivity known in the moment
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 chemiluminescent detection 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
Reactivity of this antibody on endogenous AUX1 remains to be determined.
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.