PGR5 | Proton gradient regulation 5
AS16 3985 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana, Picea abies, Pinus sylvestris
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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.
1: 1000 - 1: 5000 (WB)
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Thylakoids were isolated according to Sirpiö et al (2011, Methods Mol Biol. 2011; 775:19-30). 3 µg/µl of total chlorophyll from Arabidopsis thaliana col-0 (1) and Arabidopsis thaliana pgr5 mutant (Munekage et al. 2002, Cell) (2) were denatured in Laemmli buffer at room temperature (15 min) were separated on 12 % SDS-PAGE mini-gels with 6 M urea and blotted for 1h to a PVDF membrane using a semi-dry transfer. Membranes were blocked with 5% milk in TBS for 1h at room temperature and incubated in antisera (1:1000 in 1 % milk/TTBS) overnight at 4°C with slow agitation, then in secondary antibody (anti-rabbit IgG HRP conjugated, AS09 602, Agrisera) 1:20 000 in 1% milk/TTBS) for 2 hours at RT with slow agitation. Blots were detected for 5 min with a chemiluminescent substrate. Exposure time was 2 min.
The pgr5 mutant has a point mutation that affects the function of the PGR5 protein, therefore a band is still seen in the western blot.
Courtesy Dr. Peter Gollan, University of Turku, Finland
This product can be sold containing proclin if requested.
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