V-PPase | vacuolar H+-pyrophosphatase
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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80.8 | 73 kDa (Arabidopsis thaliana)
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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
Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.
V-PPase (pyrophosphate-energized vacuolar membrane proton pump 1), EC=126.96.36.199, 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
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.
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