Cell Signaling Technology

Product Pathways - Neuroscience

Phospho-APP (Thr668) Antibody #3823

Applications Reactivity Sensitivity MW (kDa) Source
W IP IF-IC H (M) (R) Endogenous 100-140 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
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-APP (Thr668) Antibody detects different isoforms of endogenous amyloid β (A4) precursor protein only when phosphorylated at Thr668 (or the corresponding position on other isoforms).

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr668 of human APP695. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or nocodazole-treated, and untransfected or transfected with APP, using Phospho-APP (Thr668) Antibody (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or λ-phosphatase treated (right), using Phospho-APP (Thr668) Antibody (green), β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red) and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (blue).

Background

Amyloid β (Aβ) precursor protein (APP) is a 100-140 kDa transmembrane glycoprotein that exists as several isoforms (1). The amino acid sequence of APP contains the amyloid domain, which can be released by a two-step proteolytic cleavage (1). The extracellular deposition and accumulation of the released Aβ fragments form the main components of amyloid plaques in Alzheimer's disease (AD) (1). APP can be phosphorylated at several sites, which may affect the proteolytic processing and secretion of this protein (2-5). Phosphorylation at Thr668 (a position corresponding to the APP695 isoform) by cyclin-dependent kinase is cell-cycle dependent and peaks during G2/M phase (4). APP phosphorylated at Thr668 exists in adult rat brain and correlates with cultured neuronal differentiation (5,6).

  1. Selkoe, D.J. (1996) J Biol Chem 271, 18295-8.
  2. Caporaso, G.L. et al. (1992) Proc Natl Acad Sci USA 89, 3055-9.
  3. Hung, A.Y. and Selkoe, D.J. (1994) EMBO J 13, 534-42.
  4. Suzuki, T. et al. (1994) EMBO J 13, 1114-22.
  5. Ando, K. et al. (1999) J Neurosci 19, 4421-7.
  6. Iijima, K. et al. (2000) J Neurochem 75, 1085-91.

Application References

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

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