Anti-AtpH | ATP synthase subunit c (chloroplastic)

Algae, Cannabis sativa, Cyclotella cryptica, Glycine max,  Hordeum vulgare, Oryza sativa, Ostreococcus tauri, Physcomitrium patens, Pinus thunbergii, Pisum sativum, Phaeodactylum tricornutum, Populus alba, Thalassiosira pseudonana, Zea mays, Vitis vinifera

Application example

10 ug of chlorophyll/well of Chlamydomonas reinhardtii total cell extract (1), Chlamydomonas reinhardtii subunit gamma deletion mutant thylakoid membrane fraction (2), Arabidospsis thaliana thylakoid membrane fraction (3), Chlamydomonas reinhardtii thylakoid membrane preparation (4) were separated on 12-18% acrylamide-8M urea gel and blotted to nitrocellulose membrane. Filters were blocked 1 h with 5% dry milk in 1 x PBS and probed with anti-ATP synthase subunit c antibody (AS09 591, 1: 10 000, 1h) and secondary HRP-conjugated anti-rabbit antibody (1: 10 000, 1 h) in 1 x PBS containing 5% dry milk. All steps were performed at RT with agitation. Signal was detected with chemiluminescent detection, exposure time 30’’.

Arabidopsis membrane preparation has been done according to Lezhneva et al. (2008) A novel pathway of cytochrome c biogenesis is involved in the assembly of the cytochrome b6f complex in arabidopsis chloroplasts. J Biol. Chem., 283:24608-24616 and Chlamydomonas membranes were prepared according to Chua & Bennoun (1975) Thylakoid membrane polypeptides of Chlamydomonas reinhardtii: wild-type and mutant strains deficient in photosystem II reaction center. PNAS 72:2175-2179

Courtesy Dr. Yves Choquet, CNRS, France

F-type ATPase (ATP synthase) is the universal enzyme that synthesizes ATP from ADP and phosphate using the energy stored in a transmembrane ion gradient. Multiple copies of the c subunit build up the ring structure (in spinach a 14-mer of ~112 kDa) of the membrane bound Fo-part of the enzyme.