epsilon Subunit of Tonoplast H+ATPase (V-ATPase ) antibody

Antigen: epsilon Subunit of Tonoplast H+ATPase (V-ATPase )

Clonality: Polyclonal

Host: Rabbit

Reactivity: Pineapple (Ananas comosus), Arabidopsis thaliana, Chlamydomonas reinhardtii (C. reinhardtii), Barley (Hordeum vulgare), Tomato (Solanum lycopersicum), Easter lily (Lilium longiflorum), Ficoidaceae (Mesembryanthemum), Tobacco (Nicotiana tabacum), Petunia, Ferns, Thellungiella, Maize/Corn (Zea mays)

Application: Western Blotting (WB), Immunohistochemistry (IHC)

Catalog no.: ABIN190733

Additional Information

Immunogen: KLH-conjugated synthetic peptide chosen from subunit E of plant V-ATPaseincluding Arabidopsis thaliana At4g11150. Peptide is conserved in valuolarH+-ATPase subunit E, isoform 1 to 3 (VHA-E1).

Format: Lyophilized

Description: Plant vacuole V-ATPase is responsible for energization of transport of ions andmetabolites, 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 acytosolic catalytic sector. It is related to the FoF1 ATP synthase. Alternativeprotein names: Vacuolar proton pump subunit E, Protein EMBRYO DEFECTIVE2448

Specificity: Predicted reactivity: dicots including Phaseolus sp. , Ricinus communis, Vitis vinifera and monocotsincluding: Oryza sativa, Zea mays, algae, Physcomitrella patens, bull frog, chicken, bovine, Drosophila melanogaster,human, mouse, rat.

Application Details

Application Notes: Recommended Dilution 1 : 2 000 - 1: 5000 with alkaline phosphatase (WB), 1:50 (IHC).

Purity: Serum

Storage: store lyophilized/reconstituted at -20°C, once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior toopening them to avoid any losses that might occur from lyophilized materialadhering to the cap or sides of the tubes.

Research Area: Plant/Algal Physiology, Cancer

Restrictions: For Research Use only

Images

Western Blot analysis of samples from yeast whole cell extract expressing the Arabidopsis thaliana subunit E1. Primary antibodies have been used at 1: 1: 000. Secondary antibodies at 1: 3 000. The load per well was 10 ug of total protein. Samples were separated on 10 % gel in Tris glycine SDS buffer. The proteins were transferred to a membrane (Schleicher & Schuell), blocked with 5% dry milk in Tris-buffered saline before incubation for 2 hours up to over night at RT with the primary antibody. A series of washes with TTBS was followed by incubation with a secondary antibody from).

Western Blotting: 1 - molecular weight markers (Precision Plus Protein Standards, Dual Color, Bio-Rad, 10ul), 2 -Arabidopsis thaliana leaf extract (25 ug) Experimental Conditions: Protein was separated on a 12% SDS-PAGE gel at 200 V for approximately 40 min using the mini-Protein 3 cell (Bio-Rad). Protein was transferred to PVDF at 100 V for 1h using the mini-TransBlot cell (Bio-Rad). Blocked overnight at 4ºC in 5% non-fat milk dissolved in 1xPBST. Incubated with anti-V-ATPase at 1:2 000 for 2h at RT. After several washes in 1xPBST, blot was incubated with goat anti-rabbit IgG-alkaline phosphatase conjugate (Bio-Rad) at 1:3000 for 1h at RT. Blot was developed with NBT/BCIP (Molecular Probes). Courtesy: Ms Jaimie Schnell, PhD candidate

Publications

Publications: Hinton, Sennoune, Bond et al.: "Function of a subunit isoforms of the V-ATPase in pH homeostasis and in vitro invasion of MDA-MB231 human breast cancer cells." in: The Journal of biological chemistry, Vol. 284, Issue 24, pp. 16400-8, 2009 (PubMed).

Schnell, Han, Miki et al.: "Soybean peroxidase propeptides are functional signal peptides and increase the yield of a foreign protein." in: Plant cell reports, 2010 (PubMed).