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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. australis R.Br, 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, V. vinifera |  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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Data sheet Product citations Protocols Customer reviews

Product Information

Immunogen

KLH-conjugated synthetic peptide chosen from subunit E of plant V-ATPase including Arabidopsis thaliana UniProt: Q39258-1, TAIR: 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 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 Immunofluorescence (IF), Immunohistochemistry (IHC), Western blot (WB)
Recommended dilution 1: 100 (IF), 1 : 50 (IHC), 1 : 2000-1 : 5000 (WB)
Expected | apparent MW

26 | 31 kDa (Arabidopsis thaliana)

Reactivity

Confirmed reactivity Ananas comosus, Arabidopsis thaliana, Cucumis sativus, Chara australis , 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, Vitis vinifera
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


Species of your interest not listed? Contact us
Not reactive in

Avicennia sp., mangrove plants, Schizosaccharomyces pombe

Application examples

Application examples 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 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 chemiluminescent 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.

Additional information

Additional information

Cellular [compartment marker] of tonoplast membrane.
This product can be sold containing ProClin if required.

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.

Related products

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

Background

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

Product citations

Selected references Collins et al. (2020). EPSIN1 Modulates the Plasma Membrane Abundance of FLAGELLIN SENSING2 for Effective Immune Responses . Plant Physiol. 2020 Feb 24. pii: pp.01172.2019. doi: 10.1104/pp.19.01172

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