APP antibody (C-Term)

Catalog No. ABIN2855011

This independently validated Rabbit Polyclonal antibody specifically detects APP in WB, IF, IHC (p), ICC and IHC (fro). It exhibits reactivity toward Human.

Quick Overview for APP antibody (C-Term) (ABIN2855011)

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

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This APP antibody is un-conjugated

Application: Western Blotting (WB), Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunocytochemistry (ICC), Immunohistochemistry (Frozen Sections) (IHC (fro))

Product Details anti-APP Antibody

Binding Specificity: C-Term

Cross-Reactivity: Human, Mouse, Rat

Characteristics: Rabbit Polyclonal antibody to APP
APP antibody [C2C3], C-term

Purification: Purified by antigen-affinity chromatography.

Immunogen: Recombinant protein encompassing a sequence within the C-terminus region of human APP. The exact sequence is proprietary.

Isotype: IgG

Application Details

Application Notes: WB: 1:500-1:3000. ICC/IF: 1:100-1:1000. IHC-P: 1:100-1:1000. Optimal dilutions/concentrations should be determined by the researcher. Not tested in other applications.

Comment:

Positive Control: HepG2 , U87-MG , SK-N-SH , IMR32 , SK-N-AS

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104426 (Immunohistochemistry)

by: Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin

No.: #104426

Date: 05/12/2022

Antigen: APP

Lot Number: 39694

Method validated: Immunohistochemistry

Positive Control:

Adult (24 months) mouse brain

3-month-old Taconic mouse Model 1349 brain fixed in 4% paraformaldehyde

Negative Control:

We incubated slices overnight with the blocking solution only and then processed them with the secondary antibody.

Notes:

The anti-APP antibody (Amyloid beta (A4) Precursor Protein) C-Term ABIN2855011 works in IHC-P, especially at higher concentrations (1:50 and 1:100), and without the use of any antigen retrieval treatment.

Validation Images

Staining of beta-amyloid positive cells in the adult mouse brain using ABIN2855011 at 40x magnification.

Full Methods

Primary Antibody: ABIN2855011

Secondary Antibody: goat anti-rabbit AF488-conjugated antibody (Invitrogen by Thermo Fisher Scientific, A11034, lot 1971418)

Full Protocol:

• Perfuse mice with paraformaldehyde 4% in 0.1 M phosphate buffer pH 7.4 and post-fix in the same fixative for an additional 2 h at RT.
• Wash, dehydrate, and embed samples in paraffin wax.
• Wash several times with 0.01 M PBS.
• Cut intestines and brain with a microtome into 6µm sections and mount on glass slides.
• After paraffin removal, incubate sections for 1 h at RT in PBS containing 1% albumin from chicken egg white (Sigma, A5378) and 0.3% Triton-X-100 (BioRad, 161-0407, lot 00583) to block non-specific binding sites.
• Incubate sections with primary anti-APP antibody (Amyloid beta (A4) Precursor Protein) C-Term (antibodies Online, ABIN2855011, lot 39694) diluted 1:50, 1:100, and 1:500 in PBS-BSA-PLL ON at RT.
• Wash sections 3x 5 min with 0.01 M PBS.
• Incubate sections with secondary goat anti-rabbit AF488-conjugated antibody (Invitrogen by Thermo Fisher Scientific, A11034, lot 1971418) diluted 1:500 in 0.1 M PBS for 1 h at RT.
• Wash sections 3x 5 min with 0.01M PBS.
• Mount specimens in Fluoroshield (Sigma, F6182, lot MKCB0153V).
• Acquire images with a fluorescence microscope and appropriate filter settings for AF488, e.g. Leica DM 6000B fluorescence microscope equipped with a digital camera at 40x magnification.

Experimental Notes:

Antigen retrieval treatment was also tested. In this case, sections were processed for microwave antigen retrieval for 10 min(95-100 °C) in 10 mM sodium citrate buffer (pH 6.0). After 20 min of spontaneous cooling, sections were washed twice for 5 min with distilled water and for 5 min with PBS.

Handling

Format: Liquid

Concentration: 0.2 mg/mL

Buffer: 1XPBS ( pH 7), 10 % Glycerol, 0.01 % Thimerosal

Preservative: Thimerosal (Merthiolate)

Precaution of Use: This product contains Thimerosal (Merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4°C. For long-term storage, aliquot and store at -20°C or below. Avoid multiple freeze-thaw cycles.

Target Details for APP

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

Alternative Name: amyloid beta precursor protein

Background: This gene encodes a cell surface receptor and transmembrane precursor protein that is cleaved by secretases to form a number of peptides. Some of these peptides are secreted and can bind to the acetyltransferase complex APBB1/TIP60 to promote transcriptional activation, while others form the protein basis of the amyloid plaques found in the brains of patients with Alzheimer disease. Mutations in this gene have been implicated in autosomal dominant Alzheimer disease and cerebroarterial amyloidosis (cerebral amyloid angiopathy). Multiple transcript variants encoding several different isoforms have been found for this gene.

Cellular Localization: Membrane , Membrane , clathrin-coated pit

Molecular Weight: 87 kDa

Gene ID: 351

UniProt: P05067

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