APP antibody (AA 666-670)

Catalog No. ABIN197432

The Rabbit Polyclonal anti-APP antibody (ABIN197432) specifically detects APP in WB and IF. The antibody is reactive with Human, Mouse and Rat samples. Independently validated for use in Immunofluorescence.

Quick Overview for APP antibody (AA 666-670) (ABIN197432)

Target: APP (Amyloid beta (A4) Precursor Protein (APP))

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This APP antibody is un-conjugated

Application: Western Blotting (WB), Immunofluorescence (IF)

Product Details anti-APP Antibody

Binding Specificity: AA 666-670

Specificity: This antibody detects endogenous levels of total APP protein.

Purification: Affinity Chromatography using epitope-specific immunogen.

Immunogen: peptide Sequence around amino acids 666~670 (A-V-T-P-E) derived from Human APP protein.

Application Details

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

Independent Validation (IF)

Validation #029576 (Immunofluorescence)

by: Molecular Pathology Core

No.: #029576

Date: 01/11/2014

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

Handling

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.

Handling Advice: Avoid repeated freezing and thawing.

Storage: -20 °C

Storage Comment: Store the antibody (in aliquots) at -20 °C.

Target Details for APP

Target: APP (Amyloid beta (A4) Precursor Protein (APP))

Alternative Name: Amyloid beta A4 Protein / APP

Background: 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

Gene ID: 351

NCBI Accession: NP_000475

UniProt: P05067

Pathways: Caspase Cascade in Apoptosis, EGFR Signaling Pathway, Transition Metal Ion Homeostasis, Skeletal Muscle Fiber Development, Toll-Like Receptors Cascades, Feeding Behaviour