Plant/Algal cell biology / Oryza sativa

product information

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.

KLH-conjugated peptides derived fromAtpH subunit c of Arabidopsis thaliana P56760 and Chlamydomonas reinhardtii Q37304

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.

Western blot (WB)

AS08 370 | anti-ATP synthase whole enzyme

AS08 304 | anti-ATP synthase subunit alpha

AS05 085 | anti-ATP synthase subunit beta

AS08 312 | anti-ATP synthase subunit gamma antibody

to be added when available

application information

1: 10 000 with standard ECL (WB)

8 kDa (for Arabidopsis thaliana)

Arabidopsis thaliana, Chlamydomonas reinhardtii

dicots including Glycine max, Pisum sativum, Vitis vinifera, monocolts indlucing: Hordeum vulgare, Oryza sativa, Zea mays, trees: Populus alba, Pinus thunbergii, moss: Physcomitrella patens,  green algae, Ostreococcus tauri

no confirmed exceptions from predicted reactivity known in the moment

please note that increased incubation at 95ºC (20-30 min) prior to loading is recommended to break the multimeric c-mer structure, detection of partial ring structures (e.g. 5 or 6 subunits) may occur

to be added when available

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 standard ECL (GE Healthcare), 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