V-ATPase, a | Vacuolar H+ ATPase, subunit a

AS09 466 Clonality: Polyclonal Host: Rabbit Reactivity: Arabidopsis thaliana, Cucumis sativus, Oryza sativa

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Product Information

Immunogen

KLH-cougted synthetic peptide derived from Arabidopsis thaliana V-ATPase subunit a, Q9SJT7, At2g21410

Host Rabbit

Clonality Polyclonal

Purity Serum

Format Lyophilized

Quantity 100 µl

Reconstitution For reconstitution add 100 µl of sterile water.

Storage Store lyophilized/reconstituted at -20°C; once reconstituted 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 lyophilized material adhering to the cap or sides of the tubes.

Tested applications ELISA (ELISA), Western blot (WB)

Recommended dilution 1 : 8000 (ELISA), 1 : 2000 (WB)

Expected | apparent MW

93 | 100 kDa (Arabidopsis thaliana)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Cucumis sativus, Oryza sativa

Predicted reactivity Chlamydomonas reainhardtii, Physcomitrella patens, Populus balsamifera, Ricinus communis, Vitis vinifera

Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application example

western blot detection using anti-AtVHA-a antibodies

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, a (AS09 466, 1:2000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

Additional information

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 membranes are used, one µg load per well should be sufficient.

Protocol of isolation of plant vacuolar membranes can be found here.

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.

Related products

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)

AS09 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

Plant protein extraction buffer

Secondary antibodies

Background

V-ATPase subunit a is coded by VHA-A2 gene. It has hydrogen ion transmembrane transporter activity.
Alternative names: At2g21410/F3K23.17, putative vacuolar proton-ATPase subunit, V-ATPase subunit a (100 kDa subunit), VHA-a

Product citations

Selected references Xing et al. (2016). Proteome Profile of Starch Granules Purified from Rice (Oryza sativa) Endosperm. PLoS One. 2016 Dec 19;11(12):e0168467. doi: 10.1371/journal.pone.0168467.
Migocka et al. (2013). NO3 (-)/H(+) Antiport in the Tonoplast of Cucumber Root Cells Is Stimulated by Nitrate Supply: Evidence for a Reversible Nitrate-Induced Phosphorylation of Vacuolar NO3 (-)/H(+) Antiport.PLoS One. 2013 Sep 11;8(9):e73972. doi: 10.1371/journal.pone.0073972.
Fumiyoshi et al. (2005). Novel type aquaporin SIPs, are mainly localized the ER membrane and show cell-specific expression in Arabidopsis thaliana. FEBS Lett. 579: 5814-58200.
Yoshihiro et al. (2004). Zinc transporter of Arabidopsis thaliana AtMTP1 is localized to vacuolar membranes and implicated in zinc homeostasis. Plant Cell Physiol. 45: 1749-1758.