Plant/Algal cell antibodies / Conifers

product information

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

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

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.

ELISA (ELISA), western blot (WB), immunogold (IG)

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)

0.1 % sodium azide is added as preservative. For antibody re-suspending information check the tube lable.

Antibodies will detect target protein in a few  µg of a crude preparation loaded per well. If purified preparations of vacuolar and plasma membranes are used, one µg load per well should be sufficient.

Protocol of isolation of vacuolar membrane is available here.

application information

1: 8000 (ELISA), 1: 2000 with standard ECL (WB), 1: 100 (IL)

80.8 | 73 kDa (Arabidopsis thaliana)

Arabidopsis thaliana

dicots including:  Nicotiana tabacum, Ricinus communis, Vitis vinifera, Saccharum officinarum, monocots including: Horderum vulgare, Oryza sativa, Zea mays, trees: Picea sitchensis, Populus trichocarpa

no confirmed exceptions from predicted reactivity known in the moment

Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.

Diluted antibody solution can be used 2 to 3 times within one month if it  contains 0.1 % sodium azide as preservative and is stored at -20ºC to -80ºC.

Anti-V-PPase antibody has been use in immunogold assay for TEM on root sections of Arabidopsis. Obtained results are not published yet. Labelling was detected mainly on the central vacuolar membrane. Some other membranes were also stained with a small number of gold particles.

Manufactured by Operon Biotechnologies.

Yoshihiro et al. (2006). Immunochemical analysis of aquaporin isoforms in Arabidopsis suspension-cultured cells. Biosci. Biotechnol. Biochem. 70: 980-987.

application example

1 µg and 10 µg of crude membrane fraction/lane from Arabidopsis thaliana  were separated on 12 % SDS-PAGE and blotted 1h to PVDF membrane (40 min. at 10 V using BioRad semidry transfer). Filters were blocked 1h with 5 % low-fat milk powder in TBS-T (0.05% Triton X.100). Membranes were washed 5 times with TBS-T, each time in a fresh polystyrene box and probed with anti-V-PPase antibodies (AS09 465, 1:2000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.