V-ATPase | epsilon subunit of tonoplast H+ATPase (200 µl)

325 €

AS07 213 | clonality: polyclonal | host: rabbit | reactivity: higher plants includingA.thaliana, L. esculentum, L. longiflorum, Petunia sp., N. tabaccum, Z. mays and algae Ch. reinhardtii | cellular [compartment marker] of tonoplast membrane

PRODUCT INFORMATION IN PDF

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).

antibody format

rabbit

polyclonal

serum

lyophilized

quantity

200 µl

for reconstitution add 200 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

western blot, (WB) immunohistochemistry (IHC)

related products

AS09 577 | anti-V-ATPase | epsilon subunit of tonoplast H+ATPase goat antibodies

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, Chlamydomonas reinhardtii, Hordeum vulgare, Lycopersicum esculentum, Lilium longiflorum, Mesembryanthemum sp. Nicotiana tabacum, Petunia sp., Zea mays, Pteris vittata (fern),Thellungiella sp. ,
predicted reactivity		dicots including Phaseolus sp. , Ricinus communis, Vitis vinifera and monocots including: Oryza sativa, algae, Physcomitrella patens, bull frog, chicken, bovine, Drosophila melanogaster,human, mouse, rat
not reactive in		mangrove plants, sp. Avicennia
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		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

10 µg of total leaf protein extracted with PEB (AS08 300) from (1) Arabidopsis thaliana, (3) Zea mays, and (4) Hordeum vulgare together with (2) cytosolic extract from Arabidopsis thaliana leafs were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 80 min (30V) to nitrocellulose. Filter was blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-V-ATPase (AS07 213, 1:2000, 1h) and secondary anti-rabbit (1:40000, 1h) antibody (HRP conjugated, recommended secondary antibody AS09 602) in TBS-T containing 2% low fat milk powder. Antibody incubations were followed by washings in TBS-T (15, +5, +5, +5 min). All steps were performed at RT with agitation. Signal was detected with SuperSignal West Pico (Thermo Scientific) using a GenoPlex Chemi CCD (accumulated signal 10 x 30s exposure, bin 2x2).

application example

Standard IF protocol for plant material has been used including slight fixation with formaldehyde followed by washing and incubation with primary and secondary antibodies conjugated to fluorescent dyes. Green dye visualization of anti-V-ATPase antibody (Alexa 488 Molecular Probes), red – anti-tubulin antibody.

immunolocalization using anti-V-ATPase antibodies

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