beta Amyloid antibody (C-Term)

Catalog No. ABIN5508784

Product Details anti-beta Amyloid Antibody

Target: beta Amyloid (Abeta) (Amyloid beta (Abeta))

Binding Specificity: C-Term

Reactivity: Human, Mouse, Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: This beta Amyloid antibody is un-conjugated

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

Specificity: A-beta 1-40, A-beta 1-42, A-beta 1-43 and APP

Cross-Reactivity (Details): Human, mouse, rat (other species not tested)

Purification: Protein G affinity purification

Immunogen: Synthetic peptide derived from Human Abeta 1-42 protein

Clone: 9E8-1A3

Isotype: IgG1

Application Details

Application Notes: ELISA: 1:40,000, WB: 1:4,000, IF: 1:2,000

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104425 (Immunohistochemistry)

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

No.: #104425

Date: 05/12/2022

Antigen: beta Amyloid

Lot Number: 190522A

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 beta amyloid antibody (Amyloid beta) (C-Term) ABIN5508784 works in IHC-P, especially at higher concentrations (1:200 and 1:500), and without the use of any antigen retrieval treatment.

Validation Images

Staining of beta-amyloid positive cells in the adult mouse brain using ABIN5508784 at 40x magnification.

Full Methods

Primary Antibody: ABIN5508784

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-beta amyloid (C-Term) (antibodies Online, ABIN5508784, lot 190522A) diluted 1:200, 1:500, 1:1000, and 1:2000 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: 1 mg/mL

Buffer: PBS, pH 7.4

Preservative: Without preservative

Handling Advice: Avoid freezing/thaw cycles.

Storage: -20 °C

Storage Comment: Store at -20°C to -80°C.

References for anti-beta Amyloid antibody (ABIN5508784)

Ordóñez-Gutiérrez, Posado-Fernández, Ahmadvand, Lettiero, Wu, Antón, Flores, Moghimi, Wandosell: "ImmunoPEGliposome-mediated reduction of blood and brain amyloid levels in a mouse model of Alzheimer's disease is restricted to aged animals." in: Biomaterials, Vol. 112, pp. 141-152, (2016) (PubMed).

Markoutsa, Papadia, Clemente, Flores, Antimisiaris: "Anti-Aβ-MAb and dually decorated nanoliposomes: effect of Aβ1-42 peptides on interaction with hCMEC/D3 cells." in: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft für Pharmazeutische Verfahrenstechnik e.V, Vol. 81, Issue 1, pp. 49-56, (2012) (PubMed).

Le Droumaguet, Nicolas, Brambilla, Mura, Maksimenko, De Kimpe, Salvati, Zona, Airoldi, Canovi, Gobbi, Magali, La Ferla, Nicotra, Scheper, Flores, Masserini, Andrieux, Couvreur: "Versatile and efficient targeting using a single nanoparticulate platform: application to cancer and Alzheimer's disease." in: ACS nano, Vol. 6, Issue 7, pp. 5866-79, (2012) (PubMed).

Canovi, Markoutsa, Lazar, Pampalakis, Clemente, Re, Sesana, Masserini, Salmona, Duyckaerts, Flores, Gobbi, Antimisiaris: "The binding affinity of anti-Aβ1-42 MAb-decorated nanoliposomes to Aβ1-42 peptides in vitro and to amyloid deposits in post-mortem tissue." in: Biomaterials, Vol. 32, Issue 23, pp. 5489-97, (2011) (PubMed).

Target Details for beta Amyloid

Target: beta Amyloid (Abeta) (Amyloid beta (Abeta))

Alternative Name: Amyloid beta (Abeta Products)

Background: Amyloid beta (Aβ or Abeta) is a peptide of 36 - 43 amino acids derived from the cleavage of APP (Amyloid Precursor Protein) by β- and γ-secretases. It is involved in Alzheimer´s disease, among other severe neurological disorders. It has been proved that the Aβ could be a crucial target for these important diseases treatment.

Pathways: Inflammasome