Plant/Algal cell antibodies / Populus sp.

product information

V-ATPase subunit A is a catalytic subunit of V1 complex of vacuolar ATPase. This enzyme (EC=3.6.3.14) is involved in acidification process of various compartements of eucaryotic cell.  This protein is coded by VHA-A gene. Alternative names: Vacuolar proton pump subunit alpha, vacuolar H(+)-ATPase subunit A, V-ATPase 69 kDa subunit

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana V-ATPase subunit A, O23654

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)

collection of antibodies to other vacuolar membrane proteins

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 for isolation of plant vacuolar membranes can be found here.

application information

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

68.8 | 70 kDa (Arabidopsis thaliana)

Arabidopsis thaliana

dicots including: Brassica napus, Cucumis sativus, Gossypium mexicanum, Lycopersicon esculentum, Phaseolus aureus, monocots including: Hordeum vulgare, Oryza sativa, Triticum aestivum, Zea mays, trees: Populus balsamifera, moss: Physcomitrella patens, algae: Chlamydomonas reinhardtii, Ostreococcus lucimarinus

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.

Manufactured by Operon Biotechnologies.

Yoshihiro et al. (2006) Immunochemical analysis of aquaporin isoforms in Arabidopsis suspension-cultured cells. 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-ATPase subunit A antibodies (AS09 467, 1:2000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.