IKP

V-ATPase, a | Vacuolar H+ ATPase, subunit a

Product no: AS09 466

AS09 466 Clonality: Polyclonal Host: Rabbit Reactivity: Arabidopsis thaliana, Cucumis sativus, Oryza sativa

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  • Product Info
  • Immunogen:

    KLH-cougted synthetic peptide derived from Arabidopsis thaliana V-ATPase subunit a, Q9SJT7, At2g21410

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Serum
    Format: Lyophilized
    Quantity: 100 l
    Reconstitution: For reconstitution add 100 µl 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: ELISA (ELISA), Western blot (WB)
    Recommended dilution: 1 : 8000 (ELISA), 1 : 2000 (WB)
    Expected | apparent MW:

    93 | 100 kDa (Arabidopsis thaliana)

  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Cucumis sativus, Oryza sativa
    Predicted reactivity: Chlamydomonas reainhardtii, Physcomitrium patens, Populus balsamifera, Ricinus communis, Vitis vinifera
    Species of your interest not listed? Contact us
    Not reactive in: Thermotoga neapolitana
  • Application Examples
  • Application example

    western blot detection using anti-AtVHA-a antibodies

    1 µg and 10 µg of crude membrane fraction/lane from Arabidopsis thaliana were separated on 12 % SDS-PAGE and blotted 1h to PVDF membrane (40 min. at 10 V using BioRad semidry transfer). Filters were blocked 1h with 5 % low-fat milk powder in TBS-T (0.05% Triton X.100). Membranes were washed 5 times with TBS-T, each time in a fresh polystyrene box and probed with anti-V-ATPase, a (AS09 466, 1:2000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

  • Additional Information
  • Additional information:

    0.1 % sodium azide is added as preservative. For antibody re-suspending information check the tube lable.

    Antibodies will detect target protein in a few µg of a crude preparation loaded per well. If purified preparations of vacuolar membranes are used, one µg load per well should be sufficient.

    Protocol of isolation of plant vacuolar membranes can be found here.

    Additional information (application):

    Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.

    Diluted antibody solution can be used 2 to 3 times within one month if it contains 0.1 % sodium azide as preservative and is stored at -20ºC to -80ºC.

  • Background
  • Background:

    V-ATPase subunit a is coded by VHA-A2 gene. It has hydrogen ion transmembrane transporter activity.
    Alternative names: At2g21410/F3K23.17, putative vacuolar proton-ATPase subunit, V-ATPase subunit a (100 kDa subunit), VHA-a

  • Product Citations
  • Selected references: 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.
    Migocka et al. (2013). NO3 (-)/H(+) Antiport in the Tonoplast of Cucumber Root Cells Is Stimulated by Nitrate Supply: Evidence for a Reversible Nitrate-Induced Phosphorylation of Vacuolar NO3 (-)/H(+) Antiport.PLoS One. 2013 Sep 11;8(9):e73972. doi: 10.1371/journal.pone.0073972.
    Fumiyoshi et al. (2005). Novel type aquaporin SIPs, are mainly localized the ER membrane and show cell-specific expression in Arabidopsis thaliana. FEBS Lett. 579: 5814-58200.
    Yoshihiro et al. (2004). Zinc transporter of Arabidopsis thaliana AtMTP1 is localized to vacuolar membranes and implicated in zinc homeostasis. Plant Cell Physiol. 45: 1749-1758.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Posters Collection



    Method for isolation of plant vacuolar membranes

    method for vacuolar membrane isolation

    Courtesy of Dr. Masayoshi Maeshima, Laboratory of Cell Dynamics, Graduate School of Bioagricultural Sciences Nagoya University Nagoya, Japan

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