The following protocols are for free floating mouse brain sections (see Note 26 ). Initially, certain controls should be included such as pre-adsorption of the primary antibody with the antigen and omitting of the primary and secondary antibodies as well as the avidin-peroxidase complex. For mouse-on-mouse detection it is appropriate to include omission of the primary antibody in each run. For optimal immunodetection of Aβ plaques it is not necessary to pretreat the sections with heat or formic acid as is needed for human sections. Likewise, for tau staining, pretreatment is not necessary in mouse sections. However, for IAPP staining of the mouse pancreas, epitope unmasking by boiling in citrate buffer is recommended as detailed below.
Protocol detail
Optimized Immunohistochemistry for Mouse Brain
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Adsorption
Antibodies
Antibody
Antigen
Avidin
Brain
Buffer
Citrate
Epitope
Formic acid
Human
Iapp
Mouse
Pancreas
Peroxidase
Plaques
Corresponding Organization :
Other organizations : New York University
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Variable analysis
independent variables
- Presence or absence of primary antibody for Aβ plaques
- Presence or absence of primary antibody for tau
- Presence or absence of primary antibody for IAPP
- Pretreatment of sections (heat or formic acid) for Aβ plaques
- Pretreatment of sections (boiling in citrate buffer) for IAPP
- Immunodetection of Aβ plaques
- Immunodetection of tau
- Immunodetection of IAPP
- Free floating mouse brain sections
- Pre-adsorption of the primary antibody with the antigen
- Omitting the primary and secondary antibodies
- Omitting the avidin-peroxidase complex
- Omitting the primary antibody in each run for mouse-on-mouse detection
- Pre-adsorption of the primary antibody with the antigen
- Omitting the primary and secondary antibodies
- Omitting the avidin-peroxidase complex
- Omitting the primary antibody in each run for mouse-on-mouse detection
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