Cell Signaling Technology

Product Pathways - Neuroscience

Phospho-APP (Thr668) Antibody #2451

Applications Reactivity MW (kDa) Source
W IP IHC-P H M R 100 to 140 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-APP (Thr668) Antibody detects different isoforms of endogenous amyloid beta (A4) precursor protein only when phosphorylated at Thr668 (or the corresponding position on other isoforms).

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) 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 (human), 3T3L1 (mouse) or C6 (rat) cells, untreated or nocodazole-treated (1 µg/ml, 16-18 hours), using Phospho-APP (Thr668) Antibody (upper) or APP Antibody #2452 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded Alzheimer's brain, showing cytoplasmic localization, using Phospho-APP (Thr668) Antibody.

Background

Amyloid beta (A4) precursor protein (APP) is a 100-140 kDa transmembrane glycoprotein existing as several isoforms (1). The amino acid sequence of APP contains the amyloid domain (A-beta), which can be released by a two-step proteolytic cleavage (1). The extracellular deposition and accumulation of the released A-beta fragments form the main components of amyloid plaques in Alzheimer's disease (1). APP can be phosphorylated at several sites, which may affect the proteolytic processing and secretion pathway of this protein (2-5). The phosphorylation at Thr668 (at a position corresponding to the APP695 isoform) by cyclin-dependent kinase is cell cycle dependent (G2/M-phase) (4). The APP Thr668 phosphorylated form exists in adult rat brain and correlates with cultured neuronal differentiation (5,6).

  1. Selkoe, D.J. (1996) J. Biol. Chem. 271, 18295-18298.
  2. Caporaso, G.L. et al. (1992) Proc. Natl. Acad. Sci. USA 89, 3055-3059.
  3. Hung, A.Y. and Selkoe, D.J. (1994) EMBO J. 13, 534-542.
  4. Suzuki, T. et al. (1994) EMBO J. 13, 1114-1122.
  5. Ando, K. et al. (1999) J. Neurosci. 19, 4421-4427.
  6. Iijima, K.I. et al. (2000) J. Neurochem. 75, 1085-1091.

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