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V-ATPase | Epsilon subunit of tonoplast H+ATPase

AS07 213  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: higher plants including A.comosus, A.thaliana, C.sativus, C.reinhardtii, F. margarita Swingle, H.vulgare, L.esculentum, L.longiflorum, Malus x domestica Borkh. c.v. Fuji, M. truncatula, M.crystallinum, N.tabacum, N.caerulescens, O.sativa, P.hybrida cv. Mitchell, Populus sp., P.vittata, Thellungiella sp., T. aestivum, Z.mays |  Cellular [compartment marker] of tonoplast membrane

V-ATPase | Epsilon subunit of tonoplast H+ATPase in the group Antibodies for Plant/Algal / Developmental Biology / Ion metabolism at Agrisera AB (Antibodies for research) (AS07 213)

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product information
Background

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

Immunogen

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
Clone
Purity Serum
Format Lyophilized
Quantity 50 µl
Reconstitution For reconstitution add 50 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 Immunohistochemistry (IHC), Western blot (WB)
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 : 50 (IHC), 1 : 2000-1 : 5000 (WB)
Expected | apparent MW

26 | 31 kDa (Arabidopsis thaliana)

Confirmed reactivity Ananas comosus, Arabidopsis thaliana, Cucumis sativus, Chlamydomonas reinhardtii, Fortunella margarita Swingle, Hordeum vulgare, Lycopersicum esculentum, Lilium longiflorum, Malus x domestica Borkh. c.v. Fuji, Medicago truncatula, Mesembryanthemum crystallinum, Nicotiana tabacum, Noccaea caerulescens, Oryza sativa, Petunia hybrida cv. Mitchell, Populus sp., Pteris vittata (fern), Thellungiella sp., Triticum aestivum, Zea mays
Predicted reactivity

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

Not reactive in

Avicennia sp., mangrove plants, Schizosaccharomyces pombe

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 Zhu et al. (2018). A comprehensive proteomic analysis of elaioplasts from citrus fruits reveals insights into elaioplast biogenesis and function. Hortic Res. 2018 Feb 7;5:6. doi: 10.1038/s41438-017-0014-x.
Lynch et al. (2017). Multifaceted plant responses to circumvent Phe hyperaccumulation by downregulation of flux through the shikimate pathway and by vacuolar Phe sequestration. Plant J. 2017 Dec;92(5):939-950. doi: 10.1111/tpj.13730.
Nagel et al. (2017). Arabidopsis SH3P2 is an ubiquitin-binding protein that functions together with ESCRT-I and the deubiquitylating enzyme AMSH3. Proc Natl Acad Sci U S A. 2017 Aug 7. pii: 201710866. doi: 10.1073/pnas.1710866114.
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.
Xing et al. (2016). Proteome Profile of Starch Granules Purified from Rice (Oryza sativa) Endosperm. PLoS One. 2016 Dec 19;11(12):e0168467. doi: 10.1371/journal.pone.0168467.
LaMontagne et al. (2016). Isolation of Microsomal Membrane Proteins from Arabidopsis thaliana. Curr. Protoc. Plant Biol. 1:217-234. doi: 10.1002/cppb.20020.
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.
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

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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