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product information
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| background |
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The Plasma Membrane H+ATPase is a family of proteins of ca. 100 kDa that are believed to be exclusive to the plasma membranes of plants and fungi. The protein is anchored within biological membrane which creates an electrochemical gradient used as an energy source and is essential for uptake of most metabolites and plant responses to environment, for example movement of leaves. |
| immunogen |
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KLH-conjugated synthetic peptide derived from available di and monocot, fern, mosses and algal plasma membrane ATPase sequences including Arabidopsis thaliana ATPase 1 (At2g18960) and ATPase 2,3,4,6,7,8,9 of Arabidopsis thaliana and hydrogen ATPase of Chlamydomonas reinhardtii (Q9FNS3) |
| antibody format |
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rabbit |
polyclonal |
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serum |
lyophilized |
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| quantity |
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200 µl lyophilized |
for reconstitution add 200 µl of sterile water. |
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| storage |
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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 |
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western blot (WB), immunofluorescence (IF) |
| related products |
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AS07 213 | anti-V-ATPase rabbit antibody antibodies to membrane transport system recommended secondary antibody |
| additional information |
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cellular [compartment marker] for plasma membrane |
application information
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| recommended dilution |
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1: 1000 - 1: 5000 with standard ECL OR 1: 10 000 with ECL-Advance, enhanced chemiluminescence (WB), 1: 600 - 1: 1000 (IF) |
| expected | apparent MW |
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95 kDa (Arabidopsis thaliana) |
| confirmed reactivity |
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dicots including: Arabidopsis thaliana, Cucurbita moschata, Glycine max (weak), Lycopersicon esculentum, Nicotiana tabaccum, Ricinus communis, Spinacia oleracea, monocots inlcuding: Hordeum vulgare, Zea mays, trees: Populus tremula, Pteris vittata (fern), algae: Chlamydmonas reinhardtii, |
| predicted reactivity |
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dicots (including Nicotiana tabacum, Solanum tuberosum, Mesembruanthemum crystallinum); monocots (including Avena sativa, Hordeum vulgare, Oryza sativa, conifers (Pinus thunbergii), mosses (Physocomitrella patens), algae (Dunaliella spp., Ostreococcus spp.), Saccharomyces cerevisiae |
| not reactive in |
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no confirmed exceptions from predicted reactivity known in the moment |
| additional information |
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for additional Western blot detection image please refer to the article below |
| selected references |
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Wulfetange et al. (2011). The Cytokiin Receptors of Arabidopsis thaliana are Locating Mainly to the Endoplasmic Reticulum. Plant Physiol. (in press). Sánchez-Nieto et al. (2011). Kinetics of the H+-ATPase from Dry and 5-Hours-Imbibed Maize Embryos in Its Native, Solubilized, and Reconstituted Forms. Mol. Plant. Mitani et al. (2011). Isolation and functional characterization of an influx silicon transporter in two pumpkin cultivars contrasting in silicon accumulation. Plant J. 66(2):231-240. |
application example
.jpg) 5 µg of total protein from (1) Zea mays lwhole cell, extracted with Protein Extration Buffer, PEB (AS08 300), (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, recommended secondary antibody AS09 602) 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).
immunolocalization 
Plasma membrane H+ATPase localization in Arabidopsis thaliana roots.
Arabidopsis thaliana, elongation zone, H+ATPase (green). Arabidopsis thaliana roots were fixed in para-formaldehyde for 30 minutes. Tissue cleaning has been performed before immunolocalization. Anti-rabbit H+ATPase | plasma membrane primary antibody diluted in 1: 300. Co-staining with DAPI visualized nucleus (blue color). Scale bar – 50 µm. Courtesy Dr. Taras Pasternak, Freiburg University
||| For applications or usage on species others than stated as confirmed above Agrisera offers a payment-after-testing option. To learn more about this or for any questions on this product, please use LiveChat option in the left menu or contact us at support@agrisera.com
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