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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|Recommended dilution||1 : 8000 (ELISA), 1 : 2000 (WB)|
|Expected | apparent MW||
63 | 68 kDa (Ricinus communis)
|Confirmed reactivity||Acetabularia sp. Hordeum vulgare, Oryza sativa, Pinus sylvestris, Pyrus sp. , Ricinus communis|
|Predicted reactivity||Arabidopsis thaliana, Cucumis sativus, Gossypium mexicanum, Hordeum vulgare, Populus trichocarpa, Vitis vinifera|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
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.
Manufactured by Operon Biotechnologies.
|Selected references||Kawamura et al. (2000). Tissue specificity of E subunit isoforms of plant vacuolar H(+)-ATPase and existence of isotype enzymes. J.Biol. Chem. 275:6515-6522.
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.
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.
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