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Technical Data Sheet

SK6030

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Stat Phospho-Regulation
Antibody Sampler Kit
Price

$395

Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Kit Summary
The Stat phospho-regulation antibody sampler kit can be used to examine the level of phosphorylation of Stat1 and Stat5, as well as the total expression level of Stat1, Stat3, and Stat5.
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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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Background
The Stat family of proteins (Stat1-6) function both as cytoplasmic signal transducers and as activators of transcription in response to cytokines and growth factor receptors. Stat1 is expressed as two variants of 84 and 91 kDa that become phosphorylated at Tyr-701 in response to cytokines. Active Stat1 assembles into the interferon-stimulated gene factor 3 complex. Stat3 is expressed as two variants, Stat3α (92 kDa) and Stat3β (79 kDa) that differ in expression and activity. Both are activated by phosphorylation at Tyr-705, which induces dimerization, nuclear translocation and DNA binding. Stat3α transcriptional activation may be regulated by phosphorylation at Ser-727 through the MAPK pathway, while Stat3β lacks this serine site. 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 and this Stat is constitutively active in some leukemic cell types. Both Stat5A (Tyr-694) and Stat5B (Tyr-699) are independently regulated and activated in various cell types.

Background References
Fu, X.Y. et al. (1993) Cell 74:1135.
Wakao, H. et al. (1994) EMBO J. 13:2182.
Gouilleux, F. et al. (1994) EMBO J. 13:4361.
Wen, Z. et al. (1995) Cell 82:241.
Meinke, A. et al. (1996) Mol. Cell. Biol. 16:6937-6944.
Darnell, J.E. (1997) Science 277:1630.
Biethahn, S. et al. (1999) Exp. Hematol. 27:885.
Buffer and Storage
Mouse monoclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store all at –20°C. Stable for 1 year.
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