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V-ATPase, A | Vacuolar H+-ATPase subunit A

454 €

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

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AS09 467

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product information
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

Immunogen

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

Host Rabbit
Clonality Polyclonal
Clone
Purity
Ammonium sulfate purified IgG
Format
Quantity 100 µl
Reconstitution
Storage

Store at -20°C; 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 material adhering to the cap or sides of the tubes.

Tested applications
ELISA (ELISA), Western Blot (WB)
Related products

Collection of antibodies to other vacuolar membrane proteins

Plant protein extraction buffer

Secondary antibodies

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.
application information
Recommended dilution

1: 8000 (ELISA), 1: 2000 with standard ECL (WB)

Expected | apparent MW

68.8 | 70 kDa (Arabidopsis thaliana)

Confirmed reactivity Arabidopsis thaliana, Mesembryanthemum crystallinum, Nicotiana tabacum
Predicted reactivity

dicots including: Brassica napus, Cucumis sativus, Gossypium mexicanum, Solanum lycopersicon, Phaseolus aureus, monocots including: Hordeum vulgare, Oryza sativa, Triticum aestivum, Zea mays, trees: Populus balsamifera, moss: Physcomitrella patens, algae: Chlamydomonas reinhardtii, Ostreococcus lucimarinus

Not reactive in

No confirmed exceptions from predicted reactivity known in the moment

Additional information
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.
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.

Application example

 

 

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.

 

western blot detection using anti-V-ATPase antibodies

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