Product Pathways - Lymphocyte Signaling
Phospho-AML1 (Ser249) Antibody #4327
PhosphoSitePlus® protein, site, and accession data: AML1
| 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 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 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
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).
- Wang, S. et al. (1993) Mol Cell Biol 13, 3324-3339.
- Ogawa, E. et al. (1993) Proc. Natl. Acad. Sci. USA 90, 6859-6863.
- Okuda, T. et al. (1996) Cell 84, 321-30.
- Wang, Q. et al. (1996) Proc. Natl. Acad. Sci. USA 93, 3444-3449.
- North, T.E. et al. (2004) Stem Cells 22, 158-168.
- Blyth, K. et al. (2005) Nat Rev Cancer 5, 376-387.
- Tanaka, T. et al. (1996) Mol Cell Biol 16, 3967-79.
- Zhang, Y. et al. (2004) J Biol Chem 279, 53116-25.
Application References
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Companion Products
- 4334 AML1 Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
For Research Use Only. Not For Use In Diagnostic Procedures.