V-ATPase | Epsilon subunit of tonoplast H+ATPase (goat antibody)

Product no: AS09 577

AS09 577 | Clonality: Polyclonal | Host: Goat | Reactivity: Higher plants including A.thaliana, A.strigosa, N. tabacum, S. lycopersicum

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  • Product Info
  • 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: Goat
    Clonality: Polyclonal
    Purity: Serum
    Format: Lyophilized
    Quantity: 300 l
    Reconstitution: For reconstitution add 300 l of sterile water
    Storage: Store lyophilized/reconstituted at -20C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    Tested applications: Western blot (WB)
    Recommended dilution: 1 : 1000-1 : 3000 (WB)
    Expected | apparent MW:

    26 | 31 kDa (Arabidopsis thaliana)

  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Avena strigosa, Nicotiana tabacum, Solanum lycopersicum
    Predicted reactivity: Algae, Chlamydomonas reinhardtii, Hordeum vulgare, Malus domestica, Mesembryanthemum sp., Oryza sativa,  Petunia sp.,Phaseolus sp. , Physcomitrium patens,  Pteris vittata (fern), Ricinus communis, Thellungiella sp., Zea mays, Vitis vinifera      Bull frog, Chicken, Bovine, Drosophila melanogaster, Human, Mouse, Rat
    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Application example

    western blot using goat and-V-ATPase antibodies

    6 µg of total SDS-extracted protein from Avena strigosa roots (R) and leaves (L) , were separated on NuPage LDS-PAGE 4-12% gradient acrylamide gel (Invitrogen) and blotted 1h to nitrocellulose. Filters were blocked 1h with 5% low-fat milk powder in TBS and probed with anti-V-ATPase antibodies (AS09 577 , 1:2000, 1h) and secondary anti-goat (1:5000, 1 h) antibody in TBS containing 5% low fat milk powder. Antibody incubations were followed by washings in TBS-T (containing 0.05% Tween-20, 0.1% Triton X-100) . All steps were performed at RT with agitation. Blots were scanned with a Typhoon scanner.

    Courtesy Dr. Sam Mugford (JIC), UK

  • Additional Information
  • Additional information (application):

    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.

    2 hours incubation with primary antibody is recommended over over night incubation which can contribute to increased background.

  • 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: McLoughlin et al. (2012). TheSnf1-relatedproteinkinasesSnRK2.4 andSnRK2.10 areinvolved inmaintenance ofrootsystemarchitecture duringsaltstress. Plant J. June 2012.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts


    Oxygenic photosynthesis poster by prof. Govindjee and Dr. Shevela

    Z-scheme of photosynthetic electron transport by prof. Govindjee and Dr. Björn and Dr. Shevela
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