V-PPase | vacuolar H+-pyrophosphatase

Product no: AS12 1849

AS12 1849 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, C. sativus, D. bardawil, H. vulgare, N. tabacum, P. abies, V. vinifera

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  • Product Info
  • Immunogen:

    KLH-conjugated synthetic peptide derived from Arabidopsis thaliana V-PPase,UniProt P31414, TAIR AT1G15690

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Serum
    Format: Lyophilized
    Quantity: 50 l
    Reconstitution: For reconstitution add 50 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 : 8000 (ELISA), 1 : 100 (IL), 1 : 2000 (WB)
    Expected | apparent MW:

    80.8 | 73 kDa (Arabidopsis thaliana)

  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Cucumis sativus, Dunaliella bardawil, Hordeum vulgare, Nicotiana tabacum, Picea abies, Vitis vinifera
    Predicted reactivity: Oryza sativa, Ricinus communis, Populus trichocarpa, Saccharum officinarum, Zea mays, Vigna radiata
    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 anti-VPP-ase antibodies


    Different amounts (depending of the tissue) of membrane proteins from developing xylem tissue of Norway spruce were separated in a gradient (4-15 %) SDS-PAGE gel and blotted 30 min to nitrocellulose membrane using a standard semi-dry Trans-Blot ® Turbo ™ (Bio-Rad) system. Blots were blocked with 5% non-fat milk protein for 1 h at room temperature (RT) with agitation. Blot was incubated in the primary antibody V-PPase (Agrisera) at a dilution of 1: 5 000 overnight with agitation. The antibody solution was decanted and the blot was rinsed briefly, and then washed 3 times for 15 min in TBS-T at RT with agitation. Blot was incubated in the secondary antibody (anti-rabbit IgG horseradish peroxidase conjugate, from Agrisera, AS09 602) diluted to 1: 10 000 for 1 h at RT with agitation. The blot was washed as above and the presence of V-PPase detected with ECL according to the manufacturer's instructions. Exposure time was ~ 1 second.

    Courtesy of Luis Alexis Jimenez Barboza
  • Additional Information
  • Additional information: Antibodies will detect target protein in 1 g of a crude membrane preparation loaded per well, If purified preparations of vacuolar and plasma membranes are used, less than g load per well should be sufficient
    Additional information (application): Protein or membrane sample should be treated at 70C for 10 min before loading on the gel
  • Background
  • Background:

    V-PPase (pyrophosphate-energized vacuolar membrane proton pump 1), EC=3.6.1.1, acts to establish proton gradient of often higher magnitude than H+-ATPase. Is involved in auxin transport and NaCl and drought tolerance.  Alternative names: pyrophosphate-energized inorganic pyrophosphatase 1, H(+)-PPase 1, vacuolar proton pyrophosphatase 1, vacuolar proton pyrophosphatase 3

  • Product Citations
  • Selected references: Hofmann, Wienkoop & Luthje (2022) Hypoxia-Induced Aquaporins and Regulation of Redox Homeostasis by a Trans-Plasma Membrane Electron Transport System in Maize Roots. Antioxidants (Basel). 2022 Apr 25;11(5):836. doi: 10.3390/antiox11050836. PMID: 35624700; PMCID: PMC9137787.
    Prinsi et al. (2020). Root Proteomic Analysis of Two Grapevine Rootstock Genotypes Showing Different Susceptibility to Salt Stress. Int J Mol Sci. 2020 Feb 6;21(3). pii: E1076. doi: 10.3390/ijms21031076.
    Patir-Nebioglu et al. (2019). Pyrophosphate modulates plant stress responses via SUMOylation. Elife. 2019 Feb 20;8. pii: e44213. doi: 10.7554/eLife.44213.
    Migocka et al. (2015). Cucumber Metal Transport Protein CsMTP9 is a plasma membrane H+ -coupled antiporter involved in the Mn2+ and Cd2+ efflux from root cells. Plant J. 2015 Oct 20. doi: 10.1111/tpj.13056.
    Migocka et al. (2014). Molecular and biochemical properties of two P 1B2 -ATPases, CsHMA3 and CsHMA4, from cucumber. Plant Cell Environ. 2014 Sep 11. doi: 10.1111/pce.12447.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials


    Method for isolation of plant vacuolar membranes

     

    protocol for plant vacuolar membrane isolation

    Please note that vacuolar membrane accounts for 10 % of protein of the total memrbanes of growing plant tissues. Recovery of vacuolar membrane following this protocol will depend upon amount of starting material used.

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

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This product can be purchased in 3 different volumes:

AS16 ECL-S-10, 10 ml. Trial size limited to one per customer

AS16 ECL-S-100, 100 ml

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Now available only as separate products:
1 x AS16 ECL-N
1 x AS16 ECL-S

This product can be purchased in 3 different volumes:

AS16 ECL-SN-10, 10 ml. Trial size limited to one per customer

AS16 ECL-SN, 100 ml

AS16 ECL-SN-1L, 1L

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