H+ATPase | Plasma membrane H+ATPase (rabbit antibody)
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KLH-conjugated synthetic peptide exposed to cytoplasm in H+ATPase model, derived from available di and monocot, fern, mosses and algal plasma membrane ATPase sequences including Arabidopsis thaliana ATPase 1 (UniProt: P20649, TAIR: At2g18960) and ATPase 2 (UniProt: P19456 , TAIR: At4g30190), 3 (UniProt: P20431, TAIR: At5g57350), 4 (UniProt: Q9SU58, TAIR: At3g47950), 6 (UniProt: Q9SH76, TAIR: At2g07560), 7 (UniProt: Q9LY32, TAIR: At3g60330), 8 (UniProt: Q9M2A0, TAIR: At3g42640), 9 (UniProt: Q42556, TAIR: At1g80660), 11 (UniProt: Q9LV11, TAIR: At5g62670) of Arabidopsis thaliana and hydrogen ATPase of Chlamydomonas reinhardtii (Q9FNS3)
90- 95 kDa (Arabidopsis thaliana, depending upon an isoform)
Algae, Avena sativa, Dunaliella spp., Gossypium hirsutum, Hordeum vulgare, Ostreococcus spp., Pinus thunbergii, Physocomitrella patens, Mesembruanthemum crystallinum, Mortierella elongata, Ostreococcus tauri, Saccharomyces cerevisiae, Solanum tuberosum, Ulva prolifera, Ustilago maydis
20 µg of total protein from Arabidopsis thaliana (1), Hordeum vulgare (2), Zea mays (3), Nicotiana tabaccum plasma membrane fraction, 2.5 µg (4), extracted with Protein Extration Buffer, PEB (AS08 300, homogenate the tissue with 3 to 5 volumes of the homogenizing buffer), were boiled for 10 min. in 70°C and separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in blocking reagent 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: 5 000 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 1:20 000 in 2% blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with chemiluminescence 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). Exposure time was 2 min.
Plasma membrane H+ATPase localization inArabidopsis 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 and anti-rabbit IgG secondary antibody conjugated with Alexa 555. Co-staining with DAPI visualized nucleus (blue color). Scale bar – 100 µm.
Courtesy Dr. Taras Pasternak, Freiburg University, Germany
Cellular [compartment marker] for plasma membrane
Before SDS-PAGE, centrifuge your samples at room temperature at 10 000 rpm/1 min to remove any aggregates.
H+ATPase will be less abundant in mature roots and leafs and therefore detection may require use of very sensitive reagents.
This product can be sold containing ProClin if requested.
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.
Wang et al. (2020). The Arabidopsis exocyst subunits EXO70B1 and EXO70B2 regulate FLS2 homeostasis at the plasma membrane. New Phytol. 2020 Mar 2. doi: 10.1111/nph.16515.
Kuang et al. (2019). Quantitative Proteome Analysis Reveals Changes in the Protein Landscape During Grape Berry Development With a Focus on Vacuolar Transport Proteins. Front Plant Sci. 2019 May 15;10:641. doi: 10.3389/fpls.2019.00641. eCollection 2019.
Yuan et al. (2019). Phospholipidase Dδ Negatively Regulates the Function of Resistance to Pseudomonas syringae pv. Maculicola 1 (RPM1). Front Plant Sci. 2019 Jan 18;9:1991. doi: 10.3389/fpls.2018.01991.
Zhang et all. (2018). Root plasma membrane H+-ATPase is involved in low pH-inhibited nitrogen accumulation in tea plants (Camellia sinensis L.). Plant Growth Regul (2018) 86: 423.
Roth et al. (2018). A rice Serine/Threonine receptor-like kinase regulates arbuscular mycorrhizal symbiosis at the peri-arbuscular membrane. Nat Commun. 2018 Nov 8;9(1):4677. doi: 10.1038/s41467-018-06865-z.
Wang et al. (2018). Resistance protein Pit interacts with the GEF OsSPK1 to activate OsRac1 and trigger rice immunity. Proc Natl Acad Sci U S A. 2018 Nov 16. pii: 201813058. doi: 10.1073/pnas.1813058115.
Pertl-Obermeyer et al. (2018). Dissecting the subcellular membrane proteome reveals enrichment of H+ (co-)transporters and vesicle trafficking proteins in acidic zones of Chara internodal cells. PLoS One. 2018 Aug 29;13(8):e0201480. doi: 10.1371/journal.pone.0201480.
Zhang et al. (2018). Maintenance of mesophyll potassium and regulation of plasma membrane H+-ATPase are associated with physiological responses of tea plants to drought and subsequent rehydration. The Crop Journal July 2018. (Camellia sinensis)
Seguel et al. (2018). PROHIBITIN 3 forms complexes with ISOCHORISMATE SYNTHASE 1 to regulate stress-induced salicylic acid biosynthesis in Arabidopsis. Plant Physiol. Jan 2018. DOI:10.1104/pp.17.00941
Duan et al. (2017). A Lipid-Anchored NAC Transcription Factor Is Translocated into the Nucleus and Activates Glyoxalase I Expression during Drought Stress. Plant Cell. 2017 Jul;29(7):1748-1772. doi: 10.1105/tpc.17.00044. (Nicotiana benthamiana)
Nagel et al. (2017). Arabidopsis SH3P2 is an ubiquitin-binding protein that functions together with ESCRT-I and the deubiquitylating enzyme AMSH3. Proc Natl Acad Sci U S A. 2017 Aug 7. pii: 201710866. doi: 10.1073/pnas.1710866114.
Aloui et al. (2017). The plasma membrane proteome of Medicago truncatula roots as modified by arbuscular mycorrhizal symbiosis. Mycorrhiza. 2017 Jul 19. doi: 10.1007/s00572-017-0789-5.
Lomin et al. (2017). Studies of cytokinin receptor–phosphotransmitter interaction provide evidences for the initiation of cytokinin signalling in the endoplasmic reticulum. Functional Plant Biology, CSIRO Publications. (Nicotiana benthamiana, western blot)
Kovaleva et al. (2017). Regulation of Petunia Pollen Tube Growth by Phytohormones: Identification of Their Potential Targets. DOI:10.17265/2161-6256/2016.04.004. (immunolocalization)
Liao et al. (2017). Arabidopsis E3 ubiquitin ligase PLANT U-BOX13 (PUB13) regulates chitin receptor LYSIN MOTIF RECEPTOR KINASE5 (LYK5) protein abundance. New Phytol. 2017 Feb 14. doi: 10.1111/nph.14472.
LaMontagne et al. (2016). Isolation of Microsomal Membrane Proteins from Arabidopsis thaliana. Curr. Protoc. Plant Biol. 1:217-234. doi: 10.1002/cppb.20020.
Heard et al. (2015). Identification of Regulatory and Cargo Proteins of Endosomal and Secretory Pathways in Arabidopsis thaliana by Proteomic Dissection. Mol Cell Proteomics. 2015 Jul;14(7):1796-813. doi: 10.1074/mcp.M115.050286. Epub 2015 Apr 21.
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