Plasma Membrane H+Atpase (H+Atpase) antibody

Antigen: Plasma Membrane H+Atpase (H+Atpase)

Clonality: Polyclonal

Host: Rabbit

Reactivity: Arabidopsis thaliana, Tomato (Solanum lycopersicum), Tobacco (Nicotiana tabacum), Ricinus communis, Spinach (Spinacia oleracea), Barley (Hordeum vulgare), Maize/Corn (Zea mays), Populus tremula, Ferns, Chlamydomonas reinhardtii (C. reinhardtii)

Application: Western Blotting (WB), Immunofluorescence (IF)

Catalog no.: ABIN249357

Additional Information

Immunogen: KLH-conjugated synthetic peptide derived from available di and monocot, fern,mosses and algal plasma membrane ATPase sequences including Arabidopsisthaliana ATPase 1 (At2g18960) and ATPase 2,3,4,6,7,8,9 of Arabidopsis thalianaand hydrogen ATPase of Chlamydomonas reinhardtii (Q9FNS3)

Format: Lyophilized

Description: The Plasma Membrane H+ATPase is a family of proteins of ca. 100 kDa that arebelieved to be exclusive to the plasma membranes of plants andfungi.

Specificity: Glycine max (weak). Predicted reactivity: dicots (including Nicotiana tabacum, Solanum tuberosum, Mesembruanthemumcrystallinum), monocots (including Avena sativa, Hordeum vulgare, Oryza sativa,Zea mays), conifers (Pinus thunbergii), mosses (Physocomitrella patens), algae(Dunaliella spp., Ostreococcus spp.), Saccharomyces cerevisiae.

Application Details

Application Notes: Recommended Dilution 1: 1000 - 1: 5000 with standard ECL OR 1: 10 000 with ECL-Advance, enhancedchemiluminescence (WB), 1: 600 - 1: 1000 (IF).

Buffer: Affinity purified serum in PBS pH 7.4

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

Restrictions: For Research Use only

Images

5 µg of total protein from (1) Zea mays lwhole cell, extracted with Protein Extration Buffer, PEB, (2) Hordeum vulgare leaf extracted with PEB, (3) Spinacia oleracea total cell extracted with PEB, (4) Arabidopsis thaliana were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent (GE Healthcare) in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 10 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Abcam) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL Advance detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).

anti-Plasma Membrane H+Atpase (H+Atpase) antibody (Image 2)

Publications

Publications: Page, Kropat, Hamel et al.: "Two Chlamydomonas CTR copper transporters with a novel cys-met motif are localized to the plasma membrane and function in copper assimilation." in: The Plant cell, Vol. 21, Issue 3, pp. 928-43, 2009 (PubMed).

Chen, Xiao, Wu et al.: "Nitric oxide enhances salt secretion and Na(+) sequestration in a mangrove plant, Avicennia marina, through increasing the expression of H(+)-ATPase and Na(+)/H(+) antiporter under high salinity." in: Tree physiology, Vol. 30, Issue 12, pp. 1570-85, 2010 (PubMed). Further details: The protein blots were probed with a primary antibody. H + -ATPase or Na + /H + antiporter at a dilution of 1:2000 for 2 h at room temperature with agitation.

Ito, Batth, Petzold et al.: "Analysis of the Arabidopsis Cytosolic Proteome Highlights Subcellular Partitioning of Central Plant Metabolism." in: Journal of proteome research, 2010 (PubMed).

Zhang, Peck: "Simplified enrichment of plasma membrane proteins for proteomic analyses in Arabidopsis thaliana." in: Proteomics, Vol. 11, Issue 9, pp. 1780-8, 2011 (PubMed). Method employed by authors: Western Blotting (WB)