V-ATPase, a | Vacuolar H+ ATPase, subunit a
AS09 466 Clonality: Polyclonal Host: Rabbit Reactivity: Arabidopsis thaliana, Cucumis sativus, Oryza sativa
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93 | 100 kDa (Arabidopsis thaliana)
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1 µg and 10 µg of crude membrane fraction/lane from Arabidopsis thaliana were separated on 12 % SDS-PAGE and blotted 1h to PVDF membrane (40 min. at 10 V using BioRad semidry transfer). Filters were blocked 1h with 5 % low-fat milk powder in TBS-T (0.05% Triton X.100). Membranes were washed 5 times with TBS-T, each time in a fresh polystyrene box and probed with anti-V-ATPase, a (AS09 466, 1:2000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.
0.1 % sodium azide is added as preservative. For antibody re-suspending information check the tube lable.
Antibodies will detect target protein in a few µg of a crude preparation loaded per well. If purified preparations of vacuolar membranes are used, one µg load per well should be sufficient.
Protocol of isolation of plant vacuolar membranes can be found here.
Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.
Diluted antibody solution can be used 2 to 3 times within one month if it contains 0.1 % sodium azide as preservative and is stored at -20ºC to -80ºC.
V-ATPase subunit a is coded by VHA-A2 gene. It has hydrogen ion transmembrane transporter activity.
Alternative names: At2g21410/F3K23.17, putative vacuolar proton-ATPase subunit, V-ATPase subunit a (100 kDa subunit), VHA-a
Migocka et al. (2013). NO3 (-)/H(+) Antiport in the Tonoplast of Cucumber Root Cells Is Stimulated by Nitrate Supply: Evidence for a Reversible Nitrate-Induced Phosphorylation of Vacuolar NO3 (-)/H(+) Antiport.PLoS One. 2013 Sep 11;8(9):e73972. doi: 10.1371/journal.pone.0073972.
Fumiyoshi et al. (2005). Novel type aquaporin SIPs, are mainly localized the ER membrane and show cell-specific expression in Arabidopsis thaliana. FEBS Lett. 579: 5814-58200.
Yoshihiro et al. (2004). Zinc transporter of Arabidopsis thaliana AtMTP1 is localized to vacuolar membranes and implicated in zinc homeostasis. Plant Cell Physiol. 45: 1749-1758.