Human/Mouse Phospho-APP/Protease Nexin II (T668) Antibody

(1 citations)   
  • Species Reactivity
    Human, Mouse
  • 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
    Polyclonal Rabbit IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Phosphopeptide containing the human APP/Protease Nexin II T668 site
  • Formulation
    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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human and Mouse Phospho-APP/Protease Nexin II (T668) by Western Blot. 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.
Immunohistochemistry
Phospho-APP/Protease Nexin II (T668) in Human Brain. 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & 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.
Background: APP

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).

  • References:
    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.
  • Long Name:
    Amyloid Precursor Protein
  • Entrez Gene IDs:
    351 (Human); 11820 (Mouse)
  • Alternate Names:
    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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. Membrane tethering of APP c-terminal fragments is a prerequisite for T668 phosphorylation preventing nuclear sphere generation
    Cell Signal, 2016;28(11):1725-1734.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit

CTS005 3
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Anti-Rabbit HRP-AEC Cell & Tissue Staining Kit

CTS006 3
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Immunohistochemistry Reagents
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Antigen Retrieval Reagent-Universal

CTS015 10
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Antigen Retrieval Reagent-Basic

CTS013 4
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Mounting Medium

CTS011 4
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Antigen Retrieval Reagent Sampler (50 mL each)

CTS016
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Antigen Retrieval Reagent-Acidic

CTS014
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NorthernLights Guard Mounting Media

NL996
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VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
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VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
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VisUCyte Antigen Retrieval Reagent-Universal

VCTS023
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Isotype Controls
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Normal Rabbit IgG Control

Ctrl AB-105-C 43  
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Normal Rabbit IgG Control

Ctrl, CyTOF-ready MAB1050  
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Secondary Antibodies
Description Application Cat# Citations Images  

Rabbit IgG HRP-conjugated Antibody

WB HAF008 23  
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Donkey Anti-Rabbit IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL004 10
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Donkey Anti-Rabbit IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL005 2
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Rabbit IgG APC-conjugated Antibody

Flow F0111 1  
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Rabbit IgG PE-conjugated Antibody

Flow F0110 2  
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Rabbit IgG Fluorescein-conjugated Antibody

Flow F0112 5  
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Donkey Anti-Rabbit IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL006 4
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Goat Anti-Rabbit IgG Biotinylated Antibody

WB BAF008 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Rabbit IgG Antibody

WB AF008 1
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Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC003  
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Donkey Anti-Rabbit IgG (H+L) Antibody

D-301-C-ABS2
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