Three different affinity-purified anti-peptide antibodies were combined to make this product. The concentrations of each of the three antibodies was 100 µg/mL (based on absorbance at 280nm), making the total antibody concentration 300 µg/mL. The antibody preparation is at a concentration of 100 µg/mL (based on absorbance at 280 nm). The presence of antibodies against three different regions of APP increases IHC sensitivity and fluorescence intensity.
Volume: 1000 µL
Concentration: 300 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 keyhole limpet hemocyanin (KLH) conjugates of three different synthetic peptides (named #3, #4 and #5). Peptide #3 is QHF QEK VES LEQ EAA NER QQ (which corresponds to residues #433-452 of the human sequence NP_000475.1); peptide #4 is EQK DRQ HTL KHF EHV RMV DPK K (residues #501-522); peptide #5 is EEI QDE VDE LLQ KEQ NYS DD (residues #556-575). It should be noted that these three peptide sequences are 100% identical with the mouse (P12023.3) and rat (P08592.1) gene products. 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 each of the three cognate peptides using agarose affinity matrices. The eluate concentrations were then adjusted to 300 µg/mL, mixed in equimolar concentrations (100 µg/mL is the final concentration of each antibody), 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.