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General description
Amyloid beta A4 protein (UniProt P05067; also known as ABPP, Alzheimer disease amyloid protein, Amyloid precursor protein, APP, APPI, Cerebral vascular amyloid peptide, CVAP, PN-II, PreA4, Protease nexin-II) is encoded by the APP (also known as A4, AD1) gene (Gene ID 351) in human. Amyloid precursor protein (APP) is initially produced with a signal peptide sequence (a.a. 1-17), the removal of which yields the mature protein with a large extracellular portion (a.a. 18-699), followed by a transmembrane segment (a.a. 700-723) and a cytoplasmic (a.a. 724-770) tail. APP can be further processed by the α-, β-, and γ-secretases in two alternative processing pathways. In the non-amyloidogenic pathway, APP is first cleaved by the plasma membrane-localized α-secretase to generate an N-terminal extracellular sAPPα fragment (a.a. 18-687) and a membrane-bound C-terminal fragment C83 (CTFα), which can be further cleaved by γ-secretase to produce a non-toxic small peptide p3 and a cytoplasmic APP intracellular domain (AICD). In the amyloidogenic pathway, APP undergoes β-cleavage in BACE-1 (β-site APP-cleaving enzyme)-enriched endosomes to generate an N-terminal extracellular sAPPβ fragment (a.a. 18-671) and a membrane-bound C-terminal fragment C99 (CTFβ). Subsequent cleavage of C99 by γ-secretase releases the amyloid β peptides, Aβ1-42 (672-713) & Aβ1-40 (672-711), and AICD. Aβ accumulation in the cortical and hippocampal regions of the brain is a major pathological feature of Alzheimer′s disease (AD). Aβ Ser8 phosphorylation is shown to promote Aβ aggregation into oligomeric and fibrillar assemblies and to prevent Aβ proteolytic clearance by certain proteases.
~4/8 kDa (monomer/dimer) and greater than 188 kDa (oligomer) observed. 4.514/9.028 kDa (Abeta1-42 monomer/dimer), 4.330/8.660 kDa (Abeta1-40 monomer/dimer) calculated.
Immunogen
Epitope: Abeta pSer8
KLH-conjugated linear peptide corresponding to an amyloid beta peptide-derived sequence with phosphorylated Ser8.
Application
Anti-phospho-Amyloid beta (Ser8) Antibody, clone 1E4E11 is an antibody against phospho-Amyloid beta (Ser8) for use in Western Blotting, Enzyme Immunoassay (ELISA), Immunohistochemistry, Immunofluorescence.
Research Category
Neuroscience
Research Sub Category
Neurodegenerative Diseases
Western Blotting Analysis: 2 µg/mL from a representative lot detected 2.5-100 ng of Ser8-phosphorylated synthetic Aβ1-40 peptide, but not non-phosphorylated Aβ1-40 peptide (Courtesy of Dr. Kumar and Prof. Dr. Walter, Department of Neurology, University Bonn, Germany).
Western Blotting Analysis: 2 µg/mL from a representative lot detected phosphorylated oligomeric amyloid beta (pAβ) peptides in 50 µg of brain extract from an 8-month old APP/PS1KI transgenic mouse, but not in extract from an age-matched non-transgenic mouse (Courtesy of Dr. Kumar and Prof. Dr. Walter, Department of Neurology, University Bonn, Germany).
ELISA Analysis: Clone 1E4E11 hybridoma culture supernatant detected the immunogen peptide with phosphorylated Ser8, but not the corresponding non-phosphorylated peptide (Kumar, S., et al. (2013). Acta Neuropathol. 125(5):699-709).
Immunohistochemistry Analysis: A representative lot detected age-dependent phospho-amyloid beta (pAβ) immunoreactivity and localization in paraffin-embedded APP/PS1KI mouse brain sections following antigen retrieval by heat and 88% formic acid treatments (Kumar, S., et al. (2013). Acta Neuropathol. 125(5):699-709).
Immunofluorescence Analysis: A representative lot detected phospho-amyloid beta (pAβ) immunoreactivity in cortical neurons colocalized with that detected with a non-phospho-specific Aβ antibody by dual fluorescent immunohistochemistry staining of paraffin-embedded 2-month old APP/PS1KI mice cotex sections following anigen retrieval by heat and 88% formic acid treatments (Kumar, S., et al. (2013). Acta Neuropathol. 125(5):699-709).
Western Blotting Analysis: A representative lot detected monomeric, dimeric, trimeric, and oligomeric forms of synthetic Aβ1-40 and Aβ1-42 peptides with phosphorylated Ser8, but not the non-phosphorylated Aβ1-40 and Aβ1-42 peptides (Kumar, S., et al. (2013). Acta Neuropathol. 125(5):699-709).
Western Blotting Analysis: A representative lot detected monomeric, dimeric, and oligomeric forms of phosphorylated amyloid beta (pAβ) peptides in brain extracts from 6- and 12-month old APP/PS1KI transgenic mice, but not in brain extracts from age-matched non-transgenic mice (Kumar, S., et al. (2013). Acta Neuropathol. 125(5):699-709).
Note: Formic acid (88%) treatment following heat retrieval is recommended for immunohistochemical detection of aggregated intraneuronal Abeta peptides in brain sections (Kumar, S., et al. (2013). Acta Neuropathol. 125(5):699-709; Christensen, D.Z., et al. (2009). Brain Res. 1301:116-125).
Biochem/physiol Actions
Clone 1E4E11 detected Aβ1-40 and Aβ1-42 peptides with phosphorylated Ser8, but not the non-phosphorylated Aβ peptides (Kumar, S., et al. (2013). Acta Neuropathol. 125(5):699-709).
Physical form
Format: Purified
Protein G purified.
Purified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Preparation Note
Stable for 1 year at 2-8°C from date of receipt.
Analysis Note
Identity Confirmation by Isotyping Test.
Isotyping Analysis: The identity of this monoclonal antibody is confirmed by isotyping test to be IgG1κ.
Other Notes
Concentration: Please refer to lot specific datasheet.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
| biological source | mouse |
| Quality Segment | 100 |
| antibody form | purified immunoglobulin |
| antibody product type | primary antibodies |
| clone | 1E4E11, monoclonal |
| species reactivity | human |
| technique(s) | ELISA: suitable,immunofluorescence: suitable,immunohistochemistry: suitable,western blot: suitable |
| isotype | IgG1κ |
| NCBI accession no. | NP_000475 |
| UniProt accession no. | P05067 |
| target post-translational modification | phosphorylation (pSer8) |
| Gene Information | human ... APP(351) |
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