V-ATPase | Epsilon subunit of tonoplast H+ATPase-
AS09 577 | clonality: polyclonal | host: goat | reactivity: higher plants includingA.thaliana, A.strigosa, N. tabacum, S. lycopersicum
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|Recommended dilution||1 : 1000-1 : 3000 (WB)|
|Expected | apparent MW||
26 | 31 kDa (Arabidopsis thaliana)
|Confirmed reactivity||Arabidopsis thaliana, Avena strigosa, Nicotiana tabacum, Solanum lycopersicum|
|Predicted reactivity||Algae, 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, Vitis vinifera Bull frog, Chicken, Bovine, Drosophila melanogaster, Human, Mouse, Rat|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
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.
|Selected references||McLoughlin et al. (2012). TheSnf1-relatedproteinkinasesSnRK2.4 andSnRK2.10 areinvolved inmaintenance ofrootsystemarchitecture duringsaltstress. Plant J. June 2012.|
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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