V-PPase | vacuolar H+-pyrophosphatase
AS12 1849 | clonality: polyclonal | host: rabbit | reactivity: A. thaliana, C. sativus, D. bardawil, H. vulgare, N. tabacum, P. abies
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|Recommended dilution||1 : 8000 (ELISA), 1 : 100 (IL), 1 : 2000 (WB)|
|Expected | apparent MW||
80.8 | 73 kDa (Arabidopsis thaliana)
|Confirmed reactivity||Arabidopsis thaliana, Cucumis sativus, Dunaliella bardawil, Hordeum vulgare, Nicotiana tabacum, Picea abies|
|Predicted reactivity||Oryza sativa, Ricinus communis, Populus trichocarpa, Saccharum officinarum, Vitis vinifera, Zea mays, Vigna radiata|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.
|Selected references||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.
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
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