beta Amyloid anticorps (C-Term)
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- Antigène Voir toutes beta Amyloid (Abeta) Anticorps
- beta Amyloid (Abeta) (Amyloid beta (Abeta))
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Épitope
- C-Term
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Reactivité
- Humain, Souris, Rat
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Hôte
- Souris
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Clonalité
- Monoclonal
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Conjugué
- Cet anticorp beta Amyloid est non-conjugé
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Application
- Western Blotting (WB), ELISA, Immunofluorescence (IF)
- Specificité
- A-beta 1-40, A-beta 1-42, A-beta 1-43 and APP
- Réactivité croisée (Details)
- Human, mouse, rat (other species not tested)
- Purification
- Protein G affinity purification
- Immunogène
- Synthetic peptide derived from Human Abeta 1-42 protein
- Clone
- 9E8-1A3
- Isotype
- IgG1
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- Indications d'application
- ELISA: 1:40,000, WB: 1:4,000, IF: 1:2,000
- Restrictions
- For Research Use only
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- by
- Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin
- No.
- #104425
- Date
- 12.05.2022
- Antigène
- beta Amyloid
- Numéro du lot
- 190522A
- Application validée
- Immunohistochemistry
- Contrôle positif
Adult (24 months) mouse brain
3-month-old Taconic mouse Model 1349 brain fixed in 4% paraformaldehyde
- Contrôle négative
We incubated slices overnight with the blocking solution only and then processed them with the secondary antibody.
- Conclusion
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.
- Anticorps primaire
- ABIN5508784
- Anticorps secondaire
- 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.
- 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.
Validation #104425 (Immunohistochemistry)Validation ImagesProtocole - by
- Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin
- No.
- #104493
- Date
- 15.03.2023
- Antigène
- Abeta
- Numéro du lot
- 190522A
- Application validée
- Immunohistochemistry
- Contrôle positif
3-month-old Taconic mouse Model 1349 brain fixed in 4% paraformaldehyde
- Contrôle négative
One control slice for each experimental procedure processed omitting the primary antibody; overnight incubation in diluent solution only.
- Conclusion
The beta Amyloid antibody (Amyloid beta) (C-Term) ABIN5508784 works in IHC-P, especially at higher concentrations (1:200 and 1:500). Tyramide amplification strongly enhances the positive signal.
- Anticorps primaire
- ABIN5508784
- Anticorps secondaire
- poly-HRP conjugated goat anti-rabbit antibody
- 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 6 µm 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 (N-Term) (antibodies-online, ABIN5508784, lot 190522A) diluted 1:200, 1:500, 1:1000, and 1:2000 in 0.1 M PBS-BSA-PLL ON at RT.
- Wash sections 3x 5 min with 0.01 M PBS.
- Incubate sections with poly-HRP conjugated goat anti-rabbit antibody for 1 h at RT.
- Wash sections 3x 5 min with 0.01 M PBS.
- Incubate sections Tyramide working solution (for 5 sections: 100X Tyramide stock solution 5 μL, 100X H2O2 solution 5 μL, and 1X Reaction buffer 500 μL; Thermo Fisher Scientific, B40915, lot 2086865) for 10 min.
- Stop the reaction with the Reaction Stop Reagent working solution.
- 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.
- Notes
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- Format
- Liquid
- Concentration
- 1 mg/mL
- Buffer
- PBS, pH 7.4
- Agent conservateur
- Without preservative
- Conseil sur la manipulation
- Avoid freezing/thaw cycles.
- Stock
- -20 °C
- Stockage commentaire
- Store at -20°C to -80°C.
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ImmunoPEGliposome-mediated reduction of blood and brain amyloid levels in a mouse model of Alzheimer's disease is restricted to aged animals." dans: Biomaterials, Vol. 112, pp. 141-152, (2016) (PubMed).
: "Anti-Aβ-MAb and dually decorated nanoliposomes: effect of Aβ1-42 peptides on interaction with hCMEC/D3 cells." dans: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft für Pharmazeutische Verfahrenstechnik e.V, Vol. 81, Issue 1, pp. 49-56, (2012) (PubMed).
: "Versatile and efficient targeting using a single nanoparticulate platform: application to cancer and Alzheimer's disease." dans: ACS nano, Vol. 6, Issue 7, pp. 5866-79, (2012) (PubMed).
: "The binding affinity of anti-Aβ1-42 MAb-decorated nanoliposomes to Aβ1-42 peptides in vitro and to amyloid deposits in post-mortem tissue." dans: Biomaterials, Vol. 32, Issue 23, pp. 5489-97, (2011) (PubMed).
: "
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ImmunoPEGliposome-mediated reduction of blood and brain amyloid levels in a mouse model of Alzheimer's disease is restricted to aged animals." dans: Biomaterials, Vol. 112, pp. 141-152, (2016) (PubMed).
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- Antigène
- beta Amyloid (Abeta) (Amyloid beta (Abeta))
- Autre désignation
- Amyloid beta (Abeta Produits)
- Synonymes
- anticorps AAA, anticorps ABETA, anticorps ABPP, anticorps AD1, anticorps APPI, anticorps CTFgamma, anticorps CVAP, anticorps PN-II, anticorps PN2, anticorps aaa, anticorps abeta, anticorps abpp, anticorps ad1, anticorps appi, anticorps ctfgamma, anticorps cvap, anticorps pn2, anticorps Abeta, anticorps Abpp, anticorps Adap, anticorps Ag, anticorps Cvap, anticorps E030013M08Rik, anticorps betaApp, anticorps amyloid beta precursor protein, anticorps amyloid beta (A4) precursor protein, anticorps APP, anticorps app, anticorps App
- Sujet
- 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
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