| product information
| | 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 | | immunogen | | KLH-conjugated synthetic peptide derived from Arabidopsis thaliana V-PPase,UniProt P31414, TAIR AT1G15690 | | antibody format | | rabbit | polyclonal, | | serum, | lyophilized | | | quantity | | | | storage | | 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) | | 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. | |
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, Picea abies
|
| predicted reactivity | | dicots including: Nicotiana tabacum, Ricinus communis, Vitis vinifera, Saccharum officinarum, monocots including: Horderum vulgare, Oryza sativa, Zea mays, 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 | | to be added when available, antibody release in March 2012. |
application example
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
bio-equip.com
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