Plant/Algal cell antibodies / Photosynthesis / ATP synthase

product information

ATP synthase produces ATP from ADP in the presence of a proton gradient across the membrane. F-type ATPases have two components, CF(1) - the catalytic core - and CF(0) - the membrane proton channel. CF(1) has five subunits: alpha(3), beta(3), gamma(1), delta(1), epsilon(1). CF(0) has three main subunits: a, b and c. The gamma chain is believed to be important in regulating ATPase activity and the flow of protons through the CF(0) complex. Alternative name of gamma subunit is also: F-ATPase gamma subunit.

synthetic peptides derived from Arabidopsis thalina chloroplast localized ATP synthase subunit gamma chain 1 and 2 protein sequence ( At4g04640 and At1g15700) and Chlamydomonas reinhardtii ATP synthase subunit gamma protein sequence ( A8HXL8) coupled to KLH

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 antibody

AS03 030 | anti-ATP synthase subunit beta hen antibody

AS05 071 | anti-ATP synthase subunit c antibody

AS07 260 | H+ATPase | plasma membrane H+ATPase

to be added when available

application information

1: 10 000 (WB), 1: 1000 (ELISA)

35.3 | 42  (Chlamydomonas reinhardtii)

35.6 | 38 (Spinacia oleracea)

Arabidopsis thaliana, Chlamydomonas reinhardtii

Glycine max, Lens culinaris, Nicotiana tabacum, Physcomitrella patens, Pisum sativum, Populus jackii, Vitis vinifera, cyanobacteria

no confirmed exceptions from predicted reactivity known in the moment

apparent molecular weight of subunit gamma (and as general rule most of ATP synthase subunits) is quite different between Chlamydomonas (42 kDa) and higher plants (38 kDa in spinach), see figure in Lemaire et al. (1989)

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 gamma antibody (AS08 312, 1: 25 000, 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’’ and 3 min (overexposed).

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