APP
Reactivity: Human
WB, ELISA, IHC, IF
Host: Mouse
Monoclonal
5A11E5
unconjugated
Application Notes
Western Blot: 1/500-1/1000. Immunofluorescence: 1/100-1/200. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Restrictions
For Research Use only
Validation #029576
(Immunofluorescence)
by
Molecular Pathology Core
No.
#029576
Date
01/11/2014
Antigen
Lot Number
8715
Method validated
Immunofluorescence
Positive Control
Human brain
Negative Control
Human liver
Notes
Signal was detected in positive control tissue, and no signal was seen in negative control tissue.
Validation Images
Figure 1. Micrograph image of positive control (human brain FFPE tissue). APP staining appears in green.
Figure 2: micrograph image of negative control (human liver FFPE tissue) stained with APP antibody.
Figure 3: micrograph image of isotype control (rabbit IgG isotype control antibody on human brain FFPE tissue).
Figure 4: micrograph image of secondary antibody only control (no primary antibody on human brain FFPE tissue).
Full Methods
Primary Antibody
Antibody: Amyloid beta A4 precursor protein (APP)
Catalog number: ABIN197433
Lot number: 8715
Secondary Antibody
Antibody: AlexaFluor 488 goat anti-Rabbit IgG
Lot number: 702323
Full Protocol
Sections were deparaffinized and rehydrated.
Sections were heated to 98°C for 20 min in citrate buffer pH 6.0 (Biogenex, HK086-9K) for antigen retrieval and cooled down for 20 min on the bench.
Sections were blocked in 10% NGS (normal goat serum) for 20 min at room temperature.
Sections were washed x 2 in 1xTBS buffer.
Sections were incubated with primary antibody diluted 1:100 in Antibody Diluent (Invitrogen, 003218) at 4°C overnight.
Sections were washed x 2 in 1xTBS buffer.
Sections were incubated with AlexaFluor 488 Goat anti-Rabbit IgG 1:1000 for 60 min.
Sections were washed x 3 in 1xTBS buffer.
Sections were mounted with DAPI (Invitrogen, Prolong Gold antifade reagent with DAPI) and coverslipped.
Sections were photographed with a Zeiss Axioskop2 microscope
Experimental Notes
None
Concentration
1.0 mg/mL
Buffer
PBS (without Mg2+ and Ca2+), pH 7.4 containing 150 mM NaCl, 0.02 % Sodium Azide and 50 % Glycerol.
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Amyloid beta precursor protein gene (ABPP) encodes a cell surface receptor and transmembrane precursor protein that is cleaved by secretases to form a number of peptides. Multiple transcript variants encoding several different isoforms have been found for this gene. Isoform APP695 is the predominant form in neuronal tissue, isoform APP751 and isoform APP770 are widely expressed in nonneuronal cells. Isoform APP751 is the most abundant form in T lymphocytes. ABPP is expressed in all fetal tissues examined with the highest levels in brain, kidney, heart and spleen with weak expression observed in liver, ABPP is induced during neuronal differentiation. In the adult brain, highest expression of ABPP gene is found in the frontal lobe of the cortex and in the anterior perisylvian cortex opercular gyri, moderate expression in the cerebellar cortex, the posterior perisylvian cortex opercular gyri and the temporal associated cortex. Weak expression is found in the striate, extra striate and motor cortices. Mutations in ABPP have been implicated in autosomal dominant Alzheimer disease and cerebroarterial amyloidosis (cerebral amyloid angiopathy).Synonyms: ABPP, APPI, Alzheimer disease amyloid protein, Amyloid Precursor Protein, CVAP, Cerebral vascular amyloid peptide, PreA4, Protease nexin-II