PGRL1 | PGR5-like protein 1A (chloroplastic)
AS19 4311 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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Peptide used to elicit this antibody is conserved in both isoforms PGRL1A and 1B of Zea mays
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Whole leaf protein (corresponding to 3 mg fresh weight) from the Arabidopsis thaliana lines Columbia-0 (Col-0), PGR5 knock out (pgr5-1), line-2 (line-2) and PGRL1A+B knock out (pgrl1ab) was extracted and denatured with 2x Tricine sample buffer (100mM Tris/HCl, 24% (w/v) glycerol, 8% SDS and 15mM DTT) by heating for 5 min at 70°C. The samples were separated on a 10% Tris Tricine gel and blotted overnight on PVDF-membrane using capillary blotting. The membrane was blocked with 5% milk in Tris-Buffered saline and 0.1% Tween (TBS-T (50mM Tris, 150mM NaCl, 0.1% Tween, pH 7.5)) for 1h at room temperature with agitation and subsequently incubated in the primary antibody at a dilution of 1: 5000 overnight at 4°C with agitation in 1% milk in TBS-T. The antibody solution was decanted and the blot was rinsed briefly, then washed 3 times for 10 min in TBS-T at RT with agitation. The membrane was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:10 000 in 1% milk in TBS-T for 1h at RT with agitation. The blot was washed 3 times for 10 min in TBS-T, once for 5 min in TBS (50mM Tris, 150mM NaCl, pH 7.5) and developed for 5 min with chemiluminescent detection reagent according to manufactures recommendations. Exposure time was 5 seconds.
Courtesy of Jan-Ferdinand Penzler Ludwig-Maximilians-University Munich (LMU), Dario Leister Lab, Germany
Synonymes: Ferredoxin-plastoquinone reductase 1