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-PPase | vacuolar H+-pyrophosphatase in the group Plant/Algal Antibodies / Membrane Transport System / Vacuolar membrane at Agrisera AB (Antibodies for research) (AS12 1849)


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product information

V-PPase (pyrophosphate-energized vacuolar membrane proton pump 1), EC=, 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


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

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 ĩl
Reconstitution For reconstitution add 50 ĩl of sterile water.
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications Western blot (WB)
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AS09 493 | anti-TIP1;1, TIP1;2 | tonoplast intrinistic protein 1-1, 1-2 (gamma)

AS09 494 | anti-TIP2;1 (delta) | Delta-VM23

AS09 510 | anti-TIP2;1 | tonoplast intrinistic protein 2-1

AS09 510 | anti-TIP2;2 | tonoplast intrinistic protein 2-2

AS09 511 | anti-TIP2;2 | tonoplast intrinistic protein 2-2 (C-terminal)

Plant protein extraction buffer

Secondary antibodies

Additional information Antibodies will detect target protein in 1 µg of a crude membrane preparation loaded per well. If purified preparations of vacuolar and plasma membranes are used, less than µg load per well should be sufficient.

Protocol of isolation of vacuolar membrane is available here. 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. 

application information
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.
Additional information

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.

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

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