Plant/Algal cell antibodies / Conifers

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

purified native V-ATPase subunit A from Ricinus communis

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 vacuolar proteins

AS09 467 | anti-V-ATPase subunit A | vacuolar H+-ATPase

AS09 503 | anti-V-ATPase, B | vacuolar ATP synthase subunit beta

AS09 468 | anti-V-ATPase subunit c | vacuolar H+-ATPase, subunit c (16 kDa)

ASS09 497 | anti- V-ATPase subunit D |V-type proton ATPase subunit D

AS07 213 | anti-V-ATPase subunit E of tonoplast H+ATPase

AS09 499 | anti-V-ATPase subunit H |V-type proton ATPase subunit H

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 vacuolar membranes can be found here.

application information

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

63 | 68 kDa (Ricinus communis)

Acetabularia sp. Hordeum vulgare, Oryza sativa, Pinus sylvestris, Pyrus sp. , Ricinus communis

dicots including: Arabidopsis thaliana, Cucumis sativus, Gossypium mexicanum, Vitis vinifera, monocots including: Hordeum vulgare, trees: 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.

Manufactured by Operon Biotechnologies.

Kawamura et al. (2000). Tissue specificity of E subunit isoforms of plant vacuolar H(+)-ATPase and existence of isotype enzymes. J.Biol. Chem. 275:6515-6522. Nakanishi &  Maeshima (1998). Molecular cloning of vacuolar H(+)-pyrophosphatase and its developmental expression in growing hypocotyl of mung bean. Plant Physiol. 116:589-597. Smart et al. (1998). Genes involved in osmoregulation during turgor-driven cell expansion of developing cotton fibers are differentially regulated. Plant Physiol. 116:1539-1549. Matsuura-Eno et al. (1992). Mechanism of the Decline in Vacuolar H -ATPase Activity in Mung Bean Hypocotyls during Chilling. Plant Physiology 100:718-722.

application example

65 µg/lane of purified vacuolar membranes  from Vigna radiata L. (1,3) and purified V-ATPase, 7.4 µg/lane (2,4)  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 502, 1:2000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

CBB - staining with Coomasie blue - left panel. Immunoblot - western blot detectoin - right panel.