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Technical Data Sheet

FM0291

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FXYD5/Dysadherin (Extracellular region)
Mouse Monoclonal IgG1
Price
Size
Species Reactivity
MW

$295
100 μl
Hu
45 kDa

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Western blot analysis of FXYD5 protein expression. Immunoprecipitates using FM0311 FXYD5 antibody only (lane 1) or FM0311 with A431 lysate (lane 2) or A431 input only (lane 3). Human cell lysates MDA-MB-231 (lane 4) or MeWo (lane 5). The blot was probed with mouse monoclonal FXYD5 (FM0291) at 1:1000.

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Immunocytochemical labeling of FXYD5 in paraformaldehyde fixed human MDA-MB-231 cells. The cells were labeled with mouse monoclonal FXYD5 (FM0291). 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). In addition, the Na+/K+ ATPase can interact with a group of regulatory subunits, the FXYD type I membrane protein family. This family contains FXYD1-7, and each member contains the conserved F-X-Y-D motif in the trans-membrane domain. FXYD5 (Dysadherin) is expressed in a variety of cells and tissue, as well as upregulated in carcinomas. FXYD5 contains extensive O-glycosylation, and is expressed as molecules that range from 35 to 55 kDa depending on cell type. FXYD5 can reduce E-cadherin mediated cell-cell adhesions, and may be involved in carcinogenesis.


Background References
Ino, Y. et al. (2002) PNAS, U S A. 99(1):365.
Lubarski, I. et al. (2005) J Biol Chem. 280(45):37717.
Lubarski Gotliv I. (2016) Front Cell Dev Biol. 4:26.
Immunogen
Clone (M029) was generated from a proprietary antigen related to the native human FXYD5 protein expressed in A431 epidermoid carcinoma 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 M029 mouse monoclonal antibody detects a 35-55 kDa* band on SDS-PAGE immunoblots of human A431, A549, MeWo, MDA-MB-231 cell, as well as human breast, lung, and skin tissues. The antibody works for western blot, immunoprecipitation, ELISA, and immunocytochemistry, as well as detects the FXYD5 protein 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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