V-ATPase | Epsilon subunit of tonoplast H+ATPase-

AS09 577 | Clonality: Polyclonal | Host: Goat | Reactivity: higher plants includingA.thaliana, A.strigosa, N. tabacum, S. lycopersicum

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Background		Plant vacuole V-ATPase is responsible for energization of transport of ions and metabolites, and acts as well 'house-keeping' and as a stress response enzyme. V-ATPase is a multi-subunit enzyme composed of a membrane sector and a cytosolic catalytic sector. It is related to the FoF1 ATP synthase. Alternative protein names: Vacuolar proton pump subunit E, Protein EMBRYO DEFECTIVE 2448
Immunogen		KLH-conjugated synthetic peptide chosen from subunit E of plant V-ATPase including Arabidopsis thaliana At4g11150. Peptide is conserved in vacuolar H+-ATPase subunit E, isoform 1 to 3 (VHA-E1).
Host		Goat
Clonality		Polyclonal
Clone
Purity		Serum
Format		Lyophilized
Quantity		300 µl
Reconstitution		For reconstitution add 300 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		Western blot (WB)
Related products		AS07 213 \| V-ATPase \| epsilon subunit of tonoplast H+ATPase rabbit antibodies AS08 577A \| V-ATPase \| epsilon subunit of tonoplast H+ATPase goat antibodies, affinity purified collection of antibodies to membrane transport system marker antibodies for plant cellular compartments Plant protein extraction buffer Secondary antibodies
Additional information

application information
Recommended dilution		1 : 1000-1 : 3000 (WB)
Expected \| apparent MW		26 \| 31 kDa (Arabidopsis thaliana)
Confirmed reactivity		Arabidopsis thaliana, Avena strigosa, Nicotiana tabacum, Solanum lycopersicum
Predicted reactivity		Algae, Chlamydomonas reinhardtii, Hordeum vulgare, Malus domestica, Mesembryanthemum sp., Oryza sativa, Petunia sp.,Phaseolus sp. , Physcomitrella patens, Pteris vittata (fern), Ricinus communis, Thellungiella sp., Zea mays, Vitis vinifera Bull frog, Chicken, Bovine, Drosophila melanogaster, Human, Mouse, Rat
Not reactive in		No confirmed exceptions from predicted reactivity are currently known.
Additional information		V-ATPase is very sensitive for the redox of the SDS buffer. We recommend using at least 50-100 mM DTT freshly prepared before handling the sample. 2 hours incubation with primary antibody is recommended over over night incubation which can contribute to increased background.
Selected references		McLoughlin et al. (2012). TheSnf1-relatedproteinkinasesSnRK2.4 andSnRK2.10 areinvolved inmaintenance ofrootsystemarchitecture duringsaltstress. Plant J. June 2012.

application example

6 µg of total SDS-extracted protein from Avena strigosa roots (R) and leaves (L) , were separated on NuPage LDS-PAGE 4-12% gradient acrylamide gel (Invitrogen) and blotted 1h to nitrocellulose. Filters were blocked 1h with 5% low-fat milk powder in TBS and probed with anti-V-ATPase antibodies (AS09 577 , 1:2000, 1h) and secondary anti-goat (1:5000, 1 h) antibody (Alexa 647) in TBS containing 5% low fat milk powder. Antibody incubations were followed by washings in TBS-T (containing 0.05% Tween-20, 0.1% Triton X-100) . All steps were performed at RT with agitation. Blots were scanned with a Typhoon scanner.

Courtesy Dr. Sam Mugford (JIC)

western blot using goat and-V-ATPase antibodies

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