PAB | protein in chloroplast atpase biogenesis
AS15 2833 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Zea mays
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Recombinant PAB from Zea mays, GRMZM2G110258
35 kDa (without transit peptide)
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Lanes contain ~5 µg of total protein from Zea mays seedling leaves (upper image), Arabidopsis thaliana seedling leaves (lower image), or isolated chloroplast stroma. Leaf extract from homozygous pab mutants (hypomorphic alleles), and purified recombinant antigen were analyzed as controls. Proteins were separated on 12% SDS-PAGE and blotted overnight to nitrocellulose using tank transfer. Blots were blocked for 45 min at room temperature (RT) in TBST +4% dry milk with agitation. The blot was incubated in the primary antibody at a dilution of 1: 1 000 for 2h at RT with agitation. The antibody solution was decanted and the blot was rinsed twice in TBST, then washed three times for 10 min in TBST at RT with agitation. The blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:10 000 in TBST +1% dry milk for 1h at RT with agitation. The blot was washed as above and incubated for ~2 minutes in ECL reagents prior to imaging with a LiCor digital imager. Exposure time was ~30 seconds.
Courtesy of Dr. Alison Barkan, The University of Oregon, USA
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