Immunohistochemical Analysis of Alzheimer's Disease

Immunohistochemistry (IHC) was performed as described [49 (link)]. Briefly, hippocampal sections from AD patients were retrieved with 10% (v:v) formic acid for 15 min at 37°C and then blocked with 5% (w:v) BSA. Blocked sections were stained with antibodies against Aβ and ATP6V0C overnight. Antibodies 6E10 (1:1000), ATP6V0C (1:500), MAP2 (1:3000) were used to detect human Aβ, ATP6V0C, and MAP2, respectively. Secondary antibodies used in IHC assays are as follows: Alexa Fluor 488 goat anti-rabbit IgG (H + L) (Invitrogen, A11008), Alexa Fluor 594 goat anti-mouse IgG (H + L) (Invitrogen, A11005), DyLight405-AffiniPure Goat Anti-Chicken IgY (IgG) (H + L) (Jackson ImmunoResearch, 103-475-155).

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Kim S.H., Cho Y.S., Kim Y., Park J., Yoo S.M., Gwak J., Kim Y., Gwon Y., Kam T.I, & Jung Y.K. (2023). Endolysosomal impairment by binding of amyloid beta or MAPT/Tau to V-ATPase and rescue via the HYAL-CD44 axis in Alzheimer disease. Autophagy, 19(8), 2318-2337.

Publication 2023

Alexa Fluor 488 goat anti-rabbit IgG (H + L) Alexa Fluor 594 goat anti-mouse IgG (H + L)

Alexa 594 Alexa fluor 488 Anti igg Antibodies Assays Chicken Formic acid Goat Human Immunohistochemistry Map2 Mouse Patients Rabbit

Corresponding Organization : Seoul National University

Other organizations : Sungkyunkwan University, Johns Hopkins Medicine, Johns Hopkins University

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Variable analysis

independent variables

Formic acid treatment (10% v:v) for 15 min at 37°C

dependent variables

Immunohistochemistry (IHC) staining of Aβ, ATP6V0C, and MAP2

control variables

Hippocampal sections from AD patients
Blocking with 5% (w:v) BSA

positive controls

Antibodies used: 6E10 (1:1000) for human Aβ, ATP6V0C (1:500) for ATP6V0C, MAP2 (1:3000) for MAP2

negative controls

No negative controls specified

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