showForm Anti-V-ATPase subunit A | vacuolar H+-ATPase antibodies

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V-ATPase, A | Vacuolar H+-ATPase subunit A (ammonium sulfate purified IgG)

AS09 467  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: A. thaliana, M. crystallinum, N. tabacum

V-ATPase, A | Vacuolar H+-ATPase subunit A (ammonium sulfate purified IgG) in the group Antibodies for Plant/Algal  / Membrane Transport System / Vacuolar membrane at Agrisera AB (Antibodies for research) (AS09 467)

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Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana V-ATPase subunit A, O23654, At1g78900

Host Rabbit
Clonality Polyclonal
Purity Ammonium sulfate purified IgG
Format Lyophilized
Quantity 100 µl
Reconstitution For reconstitution add 100 µl of sterile water.
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications ELISA (ELISA), Western blot (WB)
Recommended dilution 1 : 8000 (ELISA), 1 : 2000 (WB)
Expected | apparent MW

68.8 | 70 kDa (Arabidopsis thaliana)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Mesembryanthemum crystallinum, Nicotiana tabacum
Predicted reactivity Chlamydomonas reinhardtii, Brassica napus, Cucumis sativus, Gossypium mexicanum, Hordeum vulgare, Oryza sativa, Ostreococcus lucimarinus, Phaseolus aureus, Populus balsamifera, Physcomitrella patens, Solanum lycopersicon, Triticum aestivum, Zea mays
Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples

Application example

western blot detection using anti-V-ATPase antibodies

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 subunit A antibodies (AS09 467, 1:2000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

Additional information

Additional information
0.1 % sodium azide is added as preservative. For antibody re-suspending information check the tube label.

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.

Protocol for 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.

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Background

Background

V-ATPase subunit A is a catalytic subunit of V1 complex of vacuolar ATPase. This enzyme (EC=3.6.3.14) 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

Product citations

Selected references Vera-Estrella et al. (2017). Cadmium and zinc activate adaptive mechanisms in Nicotiana tabacum similar to those observed in metal tolerant plants. Planta. 2017 Apr 28. doi: 10.1007/s00425-017-2700-1.
Barkla et al. (2016). Single-cell-type quantitative proteomic and ionomic analysis of epidermal bladder cells from the halophyte model plant Mesembryanthemum crystallinum to identify salt-responsive proteins. BMC Plant Biol. 2016 May 10;16(1):110. doi: 10.1186/s12870-016-0797-1.
Yoshihiro et al. (2006) Immunochemical analysis of aquaporin isoforms in Arabidopsis suspension-cultured cells. Cells. Biosci.Biotechnol. Biochem. 70: 980-987.

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