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Technical Data Sheet

OM5741

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Osteopontin (N-terminal region) M574
Mouse Monoclonal IgG1
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
60 kDa

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Western blot analysis of human recombinant osteopontin protein (lane 1 & 2) or human MDA-MB-231 cells (lane 3 & 4). The blots were probed with mouse monoclonal anti-Osteopontin (OM5741) at a dilution of 1:1000 (lane 1), 1:4000 (lane 2), 1:250 (lane 3), and 1:1000 (lane 4).

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Immunocytochemical labeling of Osteopontin in paraformaldehyde fixed and NP-40 permeabilized MCF-7 cells. The cells were labeled with mouse monoclonal anti-Osteopontin (M574). The antibody was detected using goat anti-mouse DyLight® 594.

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Application
Dilution


ELISA
1:1000


ICC
1:50


WB
1:500



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
Osteopontin (OPN) is a 34 kDa sialic acid rich member of small integrin-binding ligand N-linked glycoproteins. It is expressed in many different tissues and is post-translationally modified at multiple sites with both glycosylation and phosphorylation. The mature post-translationally modified protein is 60 kDa. OPN is involved with cell survival, proliferation, invasion, and stem like behavior. OPN can interact with CD44, bind hydroxyapatite, and activates many integrins. These interactions are important for OPN function in cell matrix formation. A higher presence of OPN has been found in a variety of cancers, leading to increased tumor growth and metastasis. In addition, OPN is involved in type I immunity through its function as a cytokine where it can enhance production of interferon-gamma and interleukin-12 and reduce production of interleukin-10.


Background References
Sodek, J. et al. (2000) Crit Rev Oral Biol Med. 11(3):279
Shevde, L. et al. (2014) Matrix Biology 37:131
Immunogen
Clone M574 was generated from a recombinant protein containing amino acid residues in the N-terminal region of human osteopontin. This sequence has high homology with rat and mouse osteopontin.
Buffer and Storage
Mouse monoclonal antibody purified with protein A 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 M574 was purified using Protein G chromatography. The antibody detects a 60 kDa* protein corresponding to the molecular mass of Osteopontin on SDS-PAGE immunoblots of human MDA-MB-231 cells and human full length recombinant Osteopontin protein.

*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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