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Technical Data Sheet

SM1481

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Stat5 (Tyr-694), phospho-specific
Mouse Monoclonal IgG1
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
92 kDa

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Western blot analysis of human A431 cells untreated (lanes 1, 3, 5, 7 & 9) or treated with EGF (100 nM) for 60 min (lanes 2, 4, 6, 8 & 10). The blots were probed with anti-Stat1 (lanes 1 & 2), anti-Stat1 (Tyr-701) (lanes 3 & 4), anti-Stat3 (lanes 5 & 6), anti-Stat5 (lanes 7 & 8), and anti-Stat5 (Tyr-694) (lanes 9 & 10).

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Immunocytochemical labeling of Stat5 in control and pervanadate-treated A431 cells. The cells were labeled with mouse monoclonal Stat5 (SM2511) or Stat5 (Tyr-694) (SM1481) antibodies, then the antibodies were detected using appropriate secondary antibody conjugated to DyLight® 594.

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Application
Dilution


ELISA
1:2000


ICC
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 stat proteins function both as cytoplasmic signal transducers and as activators of transcription. Stat5 is activated in response to a wide variety of ligands including IL-2, GM-CSF, growth hormone and prolactin. Phosphorylation at Tyr-694 is required for Stat5A activation. Stat5 has been found to be constitutively active in some leukemic cell types. Phosphorylated Stat5 is found in some endothelial cells treated with IL-3, which suggests its involvement in angiogenesis and cell motility. Both Stat5A (Tyr-694) and Stat5B (Tyr-699) are independently regulated and activated in various cell types. For instance, both isoforms are activated in response to IFNα in B cells, but only Stat5A is phosphorylated in response to IFNα in HeLa cells.


Background References
Gouilleux, F. et al. (1994) EMBO J. 13:4361.
Wakao, H. et al. (1994) EMBO J. 13:2182.
Meinke, A. et al. (1996) Mol. Cell. Biol. 16:6937-6944.
Darnell, J.E. (1997) Science 277:1630.
Immunogen
Clone (M148) was generated from a synthetic peptide (coupled to KLH) corresponding to amino acid residues around tyrosine 694 of human Stat5A. This peptide sequence is identical to tyrosine 699 in human Stat5B, and has high homology to the conserved tyrosine site in rat and mouse Stat5A/B.
Buffer and Storage
Mouse monoclonal antibody purified with affinity 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
The antibody detects a 92 kDa* band corresponding to Stat5A on SDS-PAGE immunoblots of human A431 cells treated with EGF, but does not detect this band in control 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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