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H+ATPase | Plasma membrane H+ATPase (chicken antibody)

AS13 2671 Clonality: Polyclonal Host: Chicken  Reactivity: [global antibody] for di- and monocots, conifers, ferns, mosses, green algae cellular [compartment marker] for plasma membrane

H+ATPase | Plasma membrane H+ATPase (chicken antibody) in the group Antibodies for Plant/Algal  / Global Antibodies at Agrisera AB (Antibodies for research) (AS13 2671)

DATA SHEET IN PDF

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How to cite this product:
Product name, number (Agrisera, Sweden)

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Product Information

Immunogen

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)

Host Chicken
Clonality Polyclonal
Purity Affinity purified IgY in PBS pH 7.4
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution add 50 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)
Recommended dilution 1 : 1000-1 : 5000 (WB)
Expected | apparent MW

95 kDa (Arabidopsis thaliana)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Spinacia oleracea, Zea mays
Predicted reactivity Angomonas deanei, Avena sativa, Brassica napus, Citrus limon, Coffea canephora, Cucumis sativus, Cucurbita moschata, Dunaliella spp, Eichhornia crassipes, Emiliana huxleyi, Glycine max (weak), Hordeum vulgare, Lactobacillus johnsonii, Laishamania braziliensis, Nicotiana tabacum, Oryza sativa, Solanum lycopersicon, Solanum tuberosum, Medicago truncatula, Mesembruanthemum crystallinum), Nannochloropsis gaditana CCMP526, Nepenthes alata, Nicotiana tabaccum, Nitrospira bacterium, Oryza sativa, Ostreococcus spp., Phaseolus acutifolius, Physocomitrella patens, Picea abies, Pinus thunbergii, Populus tremula, Pteris vittata, Ricinus communis, Saccharomyces cerevisiae, Solanum lycopersicum, Strigomonas culicis, Toxoplasma gondii, Triticum urartu, Trypanosoma cruzi, Zosteria marina, Vicia faba, Vigna angularis
Species of your interest not listed? Contact us
Not reactive in

Chlamydomonas reinhardtii

Application examples

Application examples

application example

western blot using anti-H+ATPase, chicken antibodies

10 µg of total protein from whole leaf extracts of Arabidopsis thaliana (1), Zea mays (2),  Spinacia oleracea (3), extracted with Protein Extration Buffer, PEB (AS08 300), 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 2% ECL 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: 2 500 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 AS10 1489) diluted to 1:25 000  for 1h at room temperature with agitation. The blots were washed as above and developed for 5 minutes according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).

Additional information

Additional information Cellular [compartment marker] for plasma membranetissue specific immunolocalization was done on paraffin emdedded samples as described here.

VERY IMPORTANT: please, do not heat up your samples over 70°C as this might cause H+ATPase to precipitate and there will be no signal on your western blot.

Related products

Related products

AS07 260 | Anti-H+ATPase | plasma membrane H+ATPase, rabbit antibodies

antibodies to membrane transport system

recommended secondary antibody

Background

Background

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.

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