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Technical Data Sheet

NM0251

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Na+/K+ ATPase β3 (Extracellular)
Mouse Monoclonal IgG2a
Price
Size
Species Reactivity
MW

$295
100 μl
Hu
40 kDa

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Western blot of native human MDA-MB-231 (lane 1), and MeWo (lane 2) cell lysates, as well as native human breast (lane 3), lung (lane 4), skin (lane 5), and brain (lane 6) tissues. The blot was probed with anti-Na+/K+ ATPase β3 (NM0251) at 1:1000.

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Immunocytochemical labeling of Na+/K+ ATPase β3 in paraformaldehyde fixed human MeWo melanoma cells. The cells were labeled with mouse monoclonal anti-Na+/K+ ATPase β3 (clone M025). The antibody was detected using goat anti-mouse Ig DyLight® 594.

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Application
Dilution


ELISA
1:2000


ICC
1:100


IP
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
The Na+/K+ ATPase is an integral membrane heterodimer belonging to the P-type ATPase family. This ion channel uses the energy derived from ATP hydrolysis to maintain membrane potential by driving Na+ export and K+ import across the plasma membrane. It is composed of a large catalytic α subunit and a membrane-spanning auxiliary β subunit. In humans, the Na+/K+ ATPase is a binary complex of an α subunit that has four isoforms (α1-α4) and a β-subunit that has three isoforms (β1, β2, β3). Na+/K+ ATPase subunit expression has been shown to be upregulated in cancers, and inhibition of Na+/K+ ATPase activity has anti-cancer effects. The β3 subunit of Na+/K+ ATPase has increased expression in human gastric cancer tissues and cell lines, and its increased expression level predicts poor patient outcome. β3 subunit knockdown significantly inhibited cell proliferation, colony-formation ability, migration, and invasion in human gastric carcinoma cell lines.


Background References
Yoshimura, SH et al. (2008) J Cell Sci. 121:2159.
Clausen, MV et al. (2017) Front Physiol. 8:371.
Li, L et al. (2017) Oncotarget. 8(48):84285.
Immunogen
Clone (M025) was generated from a proprietary antigen related to the native human Na+/K+ ATPase β3 subunit expressed in MeWo melanoma cell line.
Buffer and Storage
Mouse monoclonal antibody purified with protein G chromatography is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
Clone M025 mouse monoclonal antibody detects a 40 kDa* protein on SDS-PAGE "Native" immunoblots of human A431, LNCaP, MeWo, MDA-MB-231, and MCF7 cells. This antibody does not detect denatured Na+/K+ ATPase β3 subunit. The antibody works for western blot, immunoprecipitation, ELISA, and immunocytochemistry, as well as detects the β3 subunit on live cells.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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