PDR8 | ABC transporter G family member 36
AS09 471 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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165 |150 kDa (Arabidopsis thaliana)
5 µg of crude membrane fraction/lane from Arabidopsis thaliana (1) and the same sample incubated with a free peptide used to produce this antibody (2) were separated on 12 % SDS-PAGE and blotted 1h to PVDF membrane (40 min. at 10 V using BioRad semidry transfer). Filters were blocked 1h with 5 % low-fat milk powder in TBS-T (0.05% Triton X.100). Membranes were washed 5 times with TBS-T, each time in a fresh polystyrene box and probed with anti-AtPDR8 antibodies (AS09 471, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.
5 day old Arabidopsis thaliana seedlings were fixed with 2% PFA in PBS for 60 minutes under vacuum. Whole seedlings were processed as described in Sauer et al., 2006 using an Insitu Pro Intavis AG robot. Rabbit anti-PDR8 antibody was diluted in 1: 400 and sheep anti-rabbit Cy3 were diluted in 1:600.
The signal could be observed mainly in epidermal cells of root apical meristems displaying an preferential outer lateral/peripheral polar localisation. In some cases an inner lateral or apolar signal could be detected.
Courtesy: Dr. Lukasz Langowski, VIB dept Plant Systems Biology, University of Ghent, Belgium
0.1 % sodium azide is added as preservative. For antibody re-suspending information check the tube lable.
Antibodies will detect target protein in a few µg of a crude preparation loaded per well. If purified preparations of vacuolar and plasma membranes are used, one µg load per well should be sufficient.
Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.
Diluted antibody solution can be used 2 to 3 times within one month if it contains 0.1 % sodium azide as preservative and is stored at -20ºC to -80ºC.
Protocol for preparation of plant plasma membranes can be found under "more information" tab. Additoinal 160 kDa cross-reacting band is detected when working with chloroplast fractions.
PDR8 protein is a key factor which controls the extent of cell death in defence response. Alternative names: ABC transporter ABCG.36, AtABCG36, pleiotropic drug resistance protein 8, protein PENETRATION 3
Yoshihiro et al. (2006). Loss of AtPDR8, a plasma membrane ABC transporter of Arabidopsis thaliana, causes hypersensitive cell death upon pathogen infection. Plant Cell Physiol. 47:309-318.
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