Cell Signaling Technology

Product Pathways - Lymphocyte Signaling

Phospho-AML1 (Ser249) Antibody #4327

Applications Reactivity Sensitivity MW (kDa) Source
W IP IF-IC F H Endogenous 55 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human
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-AML1 (Ser249) Antibody detects endogenous levels of AML1 protein only when phosphorylated on Ser249.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids around Ser249 of human AML1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts of TPA treated HEL cells, untreated or treated with λ phosphatase, using Phospho-AML1 (Ser249) Antibody (upper), or AML1 Antibody #4334 (lower).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (green) or treated with λ phosphatase (blue), using Phospho-AML1 (Ser249 ) Antibody compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated or treated with λ phosphatase, using Phospho-AML1 (Ser249) Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


Background

AML1 (also known as Runx1, CBFA2, and PEBP2αB) is a member of the core binding factor (CBF) family of transcription factors (1,2). It is required for normal development of all hematopoietic lineages (3-5). AML1 forms a heterodimeric DNA binding complex with its partner protein CBFβ and regulates the expression of cellular genes by binding to promoter and enhancer elements. AML1 is commonly translocated in hematopoietic cancers: chromosomal translocations include t(8;21) AML1-ETO, t(12;21) TEL-AML, and t(8;21) AML-M2 (6). Phosphorylation of AML1 on several potential serine and threonine sites, including Ser249, is thought to occur in an Erk-dependent manner (7,8).

  1. Wang, S. et al. (1993) Mol Cell Biol 13, 3324-3339.
  2. Ogawa, E. et al. (1993) Proc. Natl. Acad. Sci. USA 90, 6859-6863.
  3. Okuda, T. et al. (1996) Cell 84, 321-30.
  4. Wang, Q. et al. (1996) Proc. Natl. Acad. Sci. USA 93, 3444-3449.
  5. North, T.E. et al. (2004) Stem Cells 22, 158-168.
  6. Blyth, K. et al. (2005) Nat Rev Cancer 5, 376-387.
  7. Tanaka, T. et al. (1996) Mol Cell Biol 16, 3967-79.
  8. Zhang, Y. et al. (2004) J Biol Chem 279, 53116-25.

Application References

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Companion Products

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

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