V-ATPase | epsilon subunit of tonoplast H+ATPase (affinty purified)
AS09 577A | clonality: polyclonal | host: goat | reactivity: higher plants includingA.thaliana, A.strigosa, M. truncatula, N. tabacum, S. lycopersicum
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1 : 1 000 with alkaline phosphatase or 1: 3000 with ECL (WB)
|Expected | apparent MW||
26 | 31 kDa (Arabidopsis thaliana)
|Confirmed reactivity||dicots including: Arabidopsis thaliana, Medicago truncatula, Nicotiana tabacum, Solanum lycopersicum, monocots including: Avena strigosa
Chlamydomonas reinhardtii, Hordeum vulgare, Malus domestica, Mesembryanthemum sp., Oryza sativa, Petunia sp.,Phaseolus sp. ,Physcomitrella patens, Pteris vittata (fern), Ricinus communis, Thellungiella sp., Zea mays, algae, Vitis vinifera
bull frog, chicken, bovine, Drosophila melanogaster,human, mouse, rat
|Not reactive in||
no confirmed exceptions from predicted reactivity known in the moment
V-ATPase is very sensitive for the redox of the SDS buffer. We recommend using at least 50-100 mM DTT freshly prepared before handling the sample.
2 hours incubation with primary antibody is recommended over over night incubation which can contribute to increased background.
to be added when available. Antibody released in November 2009
6 µg of total SDS-extracted protein from Avena strigosa roots (R) and leaves (L) , were separated on NuPage LDS-PAGE 4-12% gradient acrylamide gel (Invitrogen) and blotted 1h to nitrocellulose. Filters were blocked 1h with 5% low-fat milk powder in TBS and probed with anti-V-ATPase antibodies (AS09 577 , 1:2000, 1h) and secondary anti-goat (1:5000, 1 h) antibody (Alexa 647) in TBS containing 5% low fat milk powder. Antibody incubations were followed by washings in TBS-T (containing 0.05% Tween-20, 0.1% Triton X-100) . All steps were performed at RT with agitation. Blots were scanned with a Typhoon scanner.
Courtesy Dr. Sam Mugford (JIC)
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