Mouse cortical neurons in culture. The green staining is APP-immunoreactivity, using fluorescein-labeled goat anti-chicken IgY (Product #F-1005, Aves Labs) and rhodamine-labeled phalloidin as a counterstain. Note the APP-staining of the neurites and growth cones, and the phalloidin-staining limited to the distal growth cones and filapodia. Photomicrograph courtesy of Dr. Philip Copenhaver (OHSU).
This antibody recognizes the peptide EQK DRQ HTL KHF EHV RMV DPK K, which corresponds to residues #501-522 of the human APP gene product. This peptide sequence is also 100% conserved between human, mouse and rat homologs of APP, and the antibodies against this sequence cross-react with all three species. The antibody preparation is at a concentration of 100 µg/mL (based on absorbance at 280 nm).
Volume: 1000 µL
Concentration: 100 µg/mL
Host Species: Chicken
Species Reactivity: Human, Mouse, Rat
Applications: ICC, IHC
Protein Name / Synonyms: Amyloid-beta A4 protein (ABPP) (APPI) (Alzheimer disease amyloid protein) (Amyloid precursor protein) (Amyloid-beta precursor protein) (Cerebral vascular amyloid peptide) (CVAP) (PreA4) (Protease nexin-II) (PN-II) [Cleaved into: N-APP; Soluble APP-alpha (S-APP-alpha); Soluble APP-beta (S-APP-beta); C99 (Beta-secretase C-terminal fragment) (Beta-CTF); Amyloid-beta protein 42 (Abeta42) (Beta-APP42); Amyloid-beta protein 40 (Abeta40) (Beta-APP40); C83 (Alpha-secretase C-terminal fragment) (Alpha-CTF); P3(42); P3(40); C80; Gamma-secretase C-terminal fragment 59 (Amyloid intracellular domain 59) (AICD-59) (AID(59)) (Gamma-CTF(59)); Gamma-secretase C-terminal fragment 57 (Amyloid intracellular domain 57) (AICD-57) (AID(57)) (Gamma-CTF(57)); Gamma-secretase C-terminal fragment 50 (Amyloid intracellular domain 50) (AICD-50) (AID(50)) (Gamma-CTF(50)); C31]
Target Description: Amyloid Precursor Protein (APP, UniProt Accession Number P05067) is a 770 amino acid, single-pass transmembrane protein whose beta-amyloid proteolytic fragment can form neurotoxic extracellular accumulations in human cerebral cortex, and is widely believed to be the cause of Alzheimer's dementia. However, the normal function of APP itself is still obscure, as are potential functions of various proteolytic fragments that have been observed in human brain [Muller, U.C., Deller, T., Korte, M. (2017) Nature Reviews Neuroscience 18: 281-298)]. To better understand these functions, we have made a set of five anti-peptide antibody reagents against APP. One of the peptide sequences used is within the beta-amyloid fragment and recognizes the extracellular amyloid plaques observed in Alzheimer patients' brains (see Cat.# ABN). Three other peptide sequences used, however, are within the extracellular domain of APP outside of this domain, and are useful for identifying the APP protein itself, and various proteolytic fragments, rather than the beta-amyloid peptide and plaques.
Gene ID: APP A4 AD1
Physical State: Liquid
Production Notes: Chickens were immunized with a keyhole limpet hemocyanin (KLH) conjugate of the synthetic peptide EQK DRQ HTL KHF EHV RMV DPK K, which corresponds to residues #501-522 of the human gene product (NP_000475.1). This sequence is also conserved in the mouse and rat APP gene products, as well. After repeated injections, immune eggs were collected from the hens, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity-purified against the cognate peptide using an agarose column, the concentration of the eluates adjusted to 100 µg/mL, and the preparation was filter-sterilized.
Validation and Application Notes
IHC Dilution Range: 1:1000-1:2000
This anti-peptide antibody mixture was analyzed by immunohistochemistry (at a dilution of 1:1000) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.
Store at 4°C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). Do not freeze these antibodies unless you want to store them for longer periods of time. Note, however, that each time an antibody preparation is frozen, about half of its binding activity is lost.
Aves Labs products are intended for use as research laboratory reagents. They are not intended for use as diagnostic or therapeutic reagents in humans.