Cell Signaling Technology

Product Pathways - Metabolism

Phospho-C/EBPalpha (Thr222/226) Antibody #2844

Applications Reactivity Sensitivity MW (kDa) Source
W H M (R) Endogenous 30, 42, 45 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat
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-C/EBPalpha (Thr222/226) Antibody detects endogenous levels of C/EBPalpha only when phosphorylated at threonine 222 and 226. This antibody does not cross-react with other phosphorylated C/EBP isoforms.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr222/226 of mouse C/EBPalpha. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from U937 cells treated with either LiCl or NaCl for the indicated times, using Phopho-C/EBPalpha (Thr222/226) Antibody (upper) and C/EBPalpha antibody (lower). C/EBPalpha phosphorylation at Thr222/226 is abolished by the specific GSK3 inhibitor LiCl, but not by NaCl, indicating that phosphorylation at these sites are depends on GSK3 kinase.

Background

CCAAT/enhancer-binding proteins (C/EBPs) are a family of transcription factors that are critical for cellular differentiation, terminal function, and inflammatory response (1). Six members of the family have been characterized (C/EBPα, β, δ, γ, ε, and ζ) and are distributed in a variety of tissues (1). Translation from alternative start codons results in two isoforms of C/EBPα (p42 and p30), which are both strong transcriptional activators (2). It has been reported that insulin and insulin-like growth factor-I stimulate the dephosphorylation of C/EBPα, which may play a key role in insulin-induced repression of GLUT4 transcription (3). Phosphorylation of C/EBPα at Thr222, Thr226, and Ser230 by GSK-3 seems to be required for adipogenesis (4).

  1. Lekstrom-Hims, J. and Xanthopoulos, K.G. (1998) J. Biol. Chem. 273, 28545-28548.
  2. Lin, F. et al. (1993) Proc. Natl. Acad. Sci. USA 90, 9606-9610.
  3. Hemati, N. et al. (1997) J. Biol. Chem. 272, 25913-25919.
  4. Ross, S.E. et al. (1999) Mol. Cell. Biol. 19, 8433-8441.

Application References

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Companion Products

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

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