V-PPase | vacuolar H+-pyrophosphatase

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AS12 1849 | clonality: polyclonal | host: rabbit | reactivity: A. thaliana, C. sativus, D. bardawil, H. vulgare, N. tabacum,  P. abies


34 st
Item No:
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

Store at -20°C; 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 material adhering to the cap or sides of the tubes.

Tested applications western blot (WB)
Related products

collection of antibodies to tonoplast proteins

AS09 492 | anti-TIP1;1, TIP1;2 | tonoplast intrinistic protein 1-1, 1-2 (gamma)

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: 2000 with standard ECL (WB), 1: 100 (IL)

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

dicots including:  Ricinus communis, Vitis vinifera, Saccharum officinarum, monocots including: Oryza sativa, Zea mays, Vigna radiata,  trees:  Populus trichocarpa

Not reactive in

no confirmed exceptions from predicted reactivity known in the moment

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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