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3 protocols using anti aβ

1

Submandibular Gland Immunohistochemistry for AD

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Paraffin-embedded submandibular gland tissue sections (10μm) from 5 male and 5 female AD and similar age controls were obtained from Banner Health (Sun City, AZ). The tissue was immunostained with anti-APP (Y188), anti-Aβ (Cell Signaling) or respective secondary only antibodies. Slides were antigen retrieved in 10mM Tris-1mM EDTA, pH 9, for 10min and rinsed with Tris-buffered saline (TBS). Antibody binding in the glands was visualized using the Vector VIP chromogen (Vector Laboratories, Burlingame, CA) reagent. Images were taken using an upright Leica DM1000 microscope and Leica DF320 digital camera system. Figures were made using Adobe Photoshop software.
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2

Comprehensive Antibody Panel for Alzheimer's Research

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anti-Aβ/APP 4G8 (mouse mAb, Covance), anti-Aβ/APP 6E10 (mouse mAb, Covance), anti-AβOs scFvA13 and scFvIm3 (ref. 13 (link)), anti-oligomer A11 (ref. 21 (link)) (rabbit pAb Millipore), anti-Aβ (rabbit mAb, Cell Signaling), anti α-Synuclein (mouse mAb, Abcam), anti-APP C-terminal fragment (rabbit pAb, Sigma), anti-APP N-terminal 22C11 (mouse mAb, Millipore), anti-V5 (mouse mAb Invitrogen, and rabbit pAb Sigma), anti-V5-HRP (mouse mAb, Sigma), anti-His tag (mouse mAb Millipore), anti-KDEL (mouse mAb, Assay Design/Stressgen) (kind gift of Prof. R. Sitia, HSR San Raffaele, Milan), anti-mTOR (rabbit pAb, Cell Signaling), anti-phospho-mTOR (rabbit pAb, Cell Signaling), anti-P70S6K (rabbit pAb, Cell Signaling) (kind gift of Dr G. Amadoro, CNR, Rome), anti-phospho-P70S6K (rabbit pAb, Cell Signaling), anti ERK1/2 (rabbit pAb, Cell Signaling), anti phosho-ERK1/2 (rabbit pAb, Cell Signaling), anti phospho-CREB (rabbit pAb, Cell Signaling), anti β-Actin (mouse mAb and rabbit pAb, Sigma), anti-calnexin (rabbit pAb, Sigma), anti-GM130 (mouse mAb, Covance), anti-Golgi 58K (mouse mAb, Sigma), anti-Rab3A (mouse mAb, Sigma), anti-Rab5 (rabbit pAb, Abcam), anti-Rab7 (goat pAb, Santa Cruz), anti-TGN46 (rabbit, pAb), anti-LAMP-1 (rabbit pAb, Abcam). anti-Rab5, anti-Rab7, anti-TGN46 are kind gift of Dr V. Triaca, CNR, Rome.
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3

Hippocampal Protein Immunoblotting

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Protein (50–80 μg) from hippocampal tissues lysed in RIPA buffer was loaded onto 10% Tris-Glycine gels and transferred to NC membranes (Millipore). Membranes were incubated with the appropriate primary antibodies (anti-Aβ, 1: 1000, Cell Signaling; GAPDH, 1: 2 500, Ab FRONTIER). GAPDH was used as the loading control. In turn, the membrane was incubated with anti-rabbit or anti-mouse secondary antibodies conjugated to horse radish peroxidase (1: 1000, Invitrogen), and are shown using an enhanced chemiluminescent substrate (Ab FRONTIER).
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