V-ATPase | Epsilon subunit of tonoplast H+ATPase

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AS07 213  clonality: polyclonal  |  host: rabbit  |  reactivity: higher plants including A.thaliana, L. esculentum L. longiflorum, O. sativa,  Petunia sp., Malus x domestica Borkh. c.v. Fuji, N. tabaccum, H.vulgare, Z. mays and algae Ch. reinhardtii |  cellular [compartment marker] of tonoplast membrane


42 st
Item No:
AS07 213

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product information

Plant vacuole V-ATPase is responsible for energization of transport of ions and metabolites, and acts as well 'house-keeping' and as a stress response enzyme. V-ATPase is a multi-subunit enzyme composed of a membrane sector and a cytosolic catalytic sector. It is related to the FoF1 ATP synthase. Alternative protein names: Vacuolar proton pump subunit E, Protein EMBRYO DEFECTIVE 2448


KLH-conjugated synthetic peptide chosen from subunit E of plant V-ATPase including Arabidopsis thaliana At4g11150. Peptide is conserved in vacuolar H+-ATPase subunit E, isoform 1 to 3 (VHA-E1).

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 ĩl
Reconstitution For reconstitution add 50 ĩl of sterile water.

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 Western blot (WB), immunohistochemistry (IHC)
Related products

AS09 577 | anti-V-ATPase | epsilon subunit of tonoplast H+ATPase goat antibodies
AS07 213P | V-ATPase | epsilon subunit of tonoplast H+ATPase | Blocking peptide 

other antibodies to vacuolar membrane

marker antibodies for plant cellular compartments

recommended secondary antibody

Additional information

cellular [compartment marker] of tonoplast membrane

application information
Recommended dilution

1 : 2 000 - 1: 5000 with alkaline phosphatase or ECL (WB), 1:50 (IHC)

Expected | apparent MW

26 | 31 kDa (Arabidopsis thaliana)

Confirmed reactivity

Ananas comosus, Arabidopsis thaliana, Cucumis sativus, Chlamydomonas reinhardtii, Hordeum vulgare, Lycopersicum esculentum, Lilium longiflorum, Malus x domestica Borkh. c.v. Fuji, Mesembryanthemum sp. Nicotiana tabacum, Noccaea caerulescens, Oryza sativa, Petunia sp., Pteris vittata (fern), Thellungiella sp., Triticum aestivum, Zea mays

Predicted reactivity

Phaseolus sp. , Physcomitrella patens, Ricinus communis, Vitis vinifera, Glyxine max, Glycine soja, Riticum aestivum, Theobroma cacao, Sorghum bicolor, Capsella rubella, Eucalypsus grandis, Citrus clementina, Citrus unshiu, Citrus limon, Solanum lycopersicum, Solanum tuberosum, Lotus japonicus, Brachypodium dystachyon, Populus trichocarpa, Prunus persica,    

bull frog, chicken, bovine, Drosophila melanogaster,human, mouse, rat

Not reactive in

mangrove plants, Avicennia sp.

Additional information

V-ATPase is very sensitive for the redox of the SDS buffer. We recommend using at least 50-100 mM DTT freshly prepared before handling the sample.

Immunostaining protocol using V-ATPase antibodies can be found here.

Selected references Liu et al. (2016). iTRAQ-based quantitative proteomic analysis reveals the role of the tonoplast in fruit senescence. J Proteomics. 2016 Sep 2;146:80-9. doi: 10.1016/j.jprot.2016.06.031.
Wattelet-Boyer et al. (2016). Enrichment of hydroxylated C24- and C26-acyl- chain sphingolipids mediates PIN2 apical sorting at trans-Golgi network subdomains. Nat Commun. 2016 Sep 29;7:12788. doi: 10.1038/ncomms12788.
Jiskrová et al. (2016). Extra- and intracellular distribution of cytokinins in the leaves of monocots and dicots. N Biotechnol. 2016 Jan 8. pii: S1871-6784(16)00002-9. doi: 10.1016/j.nbt.2015.12.010
Bancel et al. (2015). Proteomic Approach to Identify Nuclear Proteins in Wheat Grain. J Proteome Res. 2015 Sep 8.
Milner et al. (2014). Root and shoot transcriptome analysis of two ecotypes of Noccaea caerulescens uncovers the role of NcNramp1 in Cd hyperaccumulation. Plant J. 2014 Feb 18. doi: 10.1111/tpj.12480.
Shen et al. (2014). The fronds tonoplast quantitative proteomic analysis in arsenic hyperaccumulator Pteris vittata L. J Proteomics. 2014 Feb 4. pii: S1874-3919(14)00047-5. doi: 10.1016/j.jprot.2014.01.029.
Ranocha et al. (2013).Arabidopsis WAT1 is a vacuolar auxin transport facilitator required for auxin homoeostasis. Nat Commun. 2013;4:2625. doi: 10.1038/ncomms3625.
Chen et al. (2013). Mn tolerance in rice is mediated by MTP8.1, a member of the cation diffusion facilitator family. J Exp. Bot. August 20.
McLoughlin et al. (2013). Identification of novel candidate phosphatidic acid binding proteins involved in the salt stress response of Arabidopsis thaliana roots. Biochem J. Jan 17.
Rainteau et al. (2012). Acyl Chains of Phospholipase D Transphosphatidylation Products in Arabidopsis Cells: A Study Using Multiple Reaction Monitoring Mass Spectrometry. PLOS ONE (Arabidopsis thaliana suspension cells).
Lang, E.G.E., S.J. Mueller, S.N.W. Hoernstein, J. Porankiewicz-Asplund, M. Vervliet-Scheebaum, R. Reski (2010). Simultaneous isolation of pure and intact chloroplasts and mitochondria from moss as basis for sub-cellular proteomics. Plant Cell Reports, DOI: 10.1007/s00299-010-0935-4. (open source).

application example

western blot using plant anti-V-ATPase antibodies

10 µg of total protein from samples such as Arabidopsis thaliana leaf (1) , Hordeum vulgare leaf (2), Zea mays leaf (3) were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70°C for 5 min and keept on ice before loading. Protein samples were separated on 4-12% Bolt Plus gels, LDS-PAGE and blotted for 70 minutes to PVDF using tank transfer. Blots were blocked immediately following transfer in 2% blocking reagent (GE RPN 2125; Healthcare) or 5% non-fat milk dissolved in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 5 000 (in blocking reagent) for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS09 602, Agrisera) diluted to 1:25 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with TMA-6 (Lumigen) detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (VersaDoc MP 4000) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.

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