Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

Phospho-Jak2 (Tyr1008) (D4A8) Rabbit mAb #8082

Applications Reactivity Sensitivity MW (kDa) Isotype
W F H M (R) (Mk) (X) (B) (Pg) Endogenous 125 Rabbit IgG

Applications Key:  W=Western Blotting  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  X=Xenopus  B=Bovine  Pg=Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-Jak2 (Tyr1008) (D4A8) Rabbit mAb recognizes endogenous levels of Jak2 protein only when phosphorylated at Tyr1008. This antibody also reacts with Jak2 when dually phosphorylated at Tyr1007 and Tyr1008. Cross-reactivity was not observed with other Jak family members by western blot.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Tyr1008 of human Jak2 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from BaF3 cells, untreated or treated with Mouse Interleukin-3 (mIL-3) #8923 (10 ng/ml, 5 min), using Phospho-Jak2 (Tyr1008) (D4A8) Rabbit mAb (upper) or total Jak2 (D2E12) XP® Rabbit mAb #3230 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from SET-2 cells, untreated or treated with the Jak2 inhibitor FLLL31 (10 μM, 4 hr), using Phospho-Jak2 (Tyr1008) (D4A8) Rabbit mAb (upper) or total Jak2 (D2E12) XP® Rabbit mAb #3230 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from LNCaP cells, untreated or treated with growth hormone (GH) (100 ng/ml, 5 min), using Phospho-Jak2 (Tyr1008) (D4A8) Rabbit mAb (upper) or total Jak2 (D2E12) XP® Rabbit mAb #3230 (lower).


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of BaF3 cells, untreated (blue) or treated with Mouse Interleukin-3 (mIL-3) #8923 (green), using Phospho-Jak2 (Tyr1008) (D4A8) Rabbit mAb.

Background

Members of the Janus family of tyrosine kinases (Jak1, Jak2, Jak3, and Tyk2) are activated by ligands binding to a number of associated cytokine receptors (1). Upon cytokine receptor activation, Jak proteins become autophosphorylated and phosphorylate their associated receptors to provide multiple binding sites for signaling proteins. These associated signaling proteins, such as Stats (2), Shc (3), insulin receptor substrates (4), and focal adhesion kinase (FAK) (5), typically contain SH2 or other phospho-tyrosine-binding domains.

Jak2 signaling is associated with a number of cytokines, growth factors, and hormones including IL-3, IL-5, IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), erythropoietin (EPO), thrombopoietin (TPO) growth hormone, prolactin, and leptin (6-13). The oncogenic potential of Jak2 has been realized though translocations and point mutations resulting in its enhanced, de-regualated kinase activity, making Jak2 a potential therapeutic target. Jak2 gene translocations resulting in fusions with the TEL (TEL-JAK2) and PCM1 (PCM1-JAK2) have been found in leukemia patients (14,15). An activating point mutation in Jak2 resulting in a valine to phenylalanine switch at position 617 (V617F) has been implicated in myeloproliferative disorders including polycythemia vera and essential thrombocythemia (16).

  1. Leonard, W.J. and O'Shea, J.J. (1998) Annu Rev Immunol 16, 293-322.
  2. Darnell, J.E. (1997) Science 277, 1630-5.
  3. VanderKuur, J. et al. (1995) J Biol Chem 270, 7587-93.
  4. Argetsinger, L.S. et al. (1995) J Biol Chem 270, 14685-92.
  5. Zhu, T. et al. (1998) J Biol Chem 273, 10682-9.
  6. Quelle, F.W. et al. (1994) Mol Cell Biol 14, 4335-41.
  7. Witthuhn, B.A. et al. (1993) Cell 74, 227-36.
  8. Tortolani, P.J. et al. (1995) Blood 85, 3444-51.
  9. Pallard, C. et al. (1995) EMBO J 14, 2847-56.
  10. Narazaki, M. et al. (1994) Proc Natl Acad Sci USA 91, 2285-9.
  11. Argetsinger, L.S. et al. (1993) Cell 74, 237-44.
  12. Rui, H. et al. (1994) J Biol Chem 269, 5364-8.
  13. Ghilardi, N. and Skoda, R.C. (1997) Mol Endocrinol 11, 393-9.
  14. Lacronique, V. et al. (1997) Science 278, 1309-12.
  15. Reiter, A. et al. (2005) Cancer Res 65, 2662-7.
  16. Kralovics, R. et al. (2005) N Engl J Med 352, 1779-90.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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