V-ATPase, A | vacuolar H+-ATPase subunit A
AS09 502 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Acetabularia sp. Hordeum vulgare, Oryza sativa, Pinus sylvestris, Pyrus sp. , Ricinus communis
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Purified native V-ATPase subunit A from Ricinus communis, B9RHV0
63 | 68 kDa (Ricinus communis)
65 µg/lane of purified vacuolar membranes from Vigna radiata L. (1,3)and purified V-ATPase, 7.4 µg/lane(2,4) 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 subunit A antibodies (AS09 502, 1:2000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.
CBB - staining with Coomasie blue - left panel. Immunoblot - western blot detectoin - right panel.
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 and plasma membranes are used, one µg load per well should be sufficient.
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 a catalytic subunit of V1 complex of vacuolar ATPase. This enzyme (EC=184.108.40.206) is involved in acidification process of various compartements of eucaryotic cell. This protein is coded by VHA-A gene. Alternative names: Vacuolar proton pump subunit alpha, vacuolar H(+)-ATPase subunit A, V-ATPase 69 kDa subunit
Nakanishi & Maeshima (1998). Molecular cloning of vacuolar H(+)-pyrophosphatase and its developmental expression in growing hypocotyl of mung bean. Plant Physiol. 116:589-597.
Smart et al. (1998). Genes involved in osmoregulation during turgor-driven cell expansion of developing cotton fibers are differentially regulated. Plant Physiol. 116:1539-1549.
Matsuura-Eno et al. (1992). Mechanism of the Decline in Vacuolar H -ATPase Activity in Mung Bean Hypocotyls during Chilling. Plant Physiology 100:718-722.
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