APP [p Thr668] Antibody

Western blot shows lysates of L‑929 mouse fibroblast cell line and HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 1 μg/mL nocodazole for 16 hours. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse Phospho-APP/Protease Nexin II (T668) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2508), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

APP/Protease Nexin II phosphorylated at T668 was detected in immersion fixed paraffin-embedded sections of human Alzheimer's brain (cortex) using Rabbit Anti-Human/Mouse Phospho-APP/Protease Nexin II (T668) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2508) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

• Reactivity: Hu, Mu
• Applications: WB, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

APP [p Thr668] Antibody Summary

• Immunogen: Phosphopeptide containing the human APP/Protease Nexin II T668 site
• Modification: p Thr668
• Specificity: Detects multiple isoforms of human and mouse APP/Protease Nexin II when phosphorylated at sites corresponding to T668 of the human APP695 isoform.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: APP
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 0.5 ug/mL
  • Immunohistochemistry 5-15 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Purity: Immunogen affinity purified
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for APP [p Thr668] Antibody

• amyloid beta (A4) precursor protein-binding, family B, member 2
• amyloid beta A4 precursor protein-binding family B member 2
• Amyloid beta precursor protein
• Amyloid beta
• APP
• beta Amyloid
• Protease Nexin II

Background

Amyloid precursor protein (APP) is a type I membrane protein with several human isoforms due to alternative splicing. APP-770, -751, and -733 contain a Kunitz protease inhibitor (KPI) domain (residue 291‑342) and APP-695 does not. APP is a cell surface molecule with many functions. It can be processed proteolytically in two different pathways. In one pathway, beta - and gamma -secretase cleave at the  beta  site between residue 670 and 671 and the gamma site between residue 711 and 714 to produce beta ‑amyloid peptide (A beta 40 and A beta 42), a major component in plaques found in brains of patients with Alzheimer's disease (1). The other pathway involves alpha -secretase that cleaves residues between 687 and 688. It is antiamyloidogenic due to its benign character and the prevention of the A beta peptide formation (2). Soluble APP containing the KPI domain, also referred to as protease nexin II, is a potent inhibitor of serine proteases and may have additional functions. For example, it may regulate the contact face of blood coagulation and limit thrombosis specially in brain due to its localization and coagulation factor XI inhibiting activity (3, 4).

1. Haass, C. (2004) EMBO J. 23:483.
2. Lichtenthaler, S. F. and C. Haass (2004) J. Clin. Invest. 113:1384.
3. Badellino, K.O. and P.N. Walsh (2000) Biochemistry 39:4769.
4. Xu, F. et al. (2005) Proc. Natl. Acad. Sci USA. 102:18135.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for APP Antibody (AF2508)(1)

Publication using AF2508

• Membrane tethering of APP c-terminal fragments is a prerequisite for T668 phosphorylation preventing nuclear sphere generation Cell Signal, 2016;28(11):1725-1734. 2016 [PMID: 27534516] (Western Blot, Human)

Bioinformatics

• Gene Symbol: APP