Immunofluorescent Staining of Mouse Brain Samples

Mouse brain samples for immunofluorescent staining were prepared as previously reported [20 (link), 21 (link)]. Sections for staining were incubated with the following primary antibodies: rabbit anti Iba1 (1:2000, Wako) and mouse anti-Aβ (6E10, 1:1000, Covance) at 4 °C overnight. After being washing with PBS, the sections were incubated with donkey anti-rabbit Alexa 488 (1:500; Jackson ImmunoResearch) and donkey anti-mouse Cy3 (1:500; Jackson ImmunoResearch) at 4 °C for 2 h. After further washing, the sections were mounted in Vectashield anti-fading medium (Vector Laboratories). The fluorescence images were observed using a confocal laser scanning microscope (CLSM; C2si, Nikon).

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Ni J., Wu Z., Meng J., Saito T., Saido T.C., Qing H, & Nakanishi H. (2019). An impaired intrinsic microglial clock system induces neuroinflammatory alterations in the early stage of amyloid precursor protein knock-in mouse brain. Journal of Neuroinflammation, 16, 173.

Publication 2019

rabbit anti Iba1 mouse anti-Aβ (6E10, Alexa 488 donkey anti-mouse Cy3 Vectashield anti-fading medium

Antibodies Brain Confocal laser scanning microscope Donkey Fluorescence Immunofluorescent Mouse Rabbit Vector

Corresponding Organization : Yasuda Women's University

Other organizations : RIKEN Center for Brain Science, Beijing Institute of Technology

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Variable analysis

independent variables

Primary antibodies used for immunofluorescent staining: rabbit anti Iba1 and mouse anti-Aβ (6E10)

dependent variables

Fluorescence images observed using a confocal laser scanning microscope (CLSM)

control variables

Mouse brain samples for immunofluorescent staining were prepared as previously reported [20, 21]
Sections were incubated with primary antibodies at 4 °C overnight
Sections were incubated with secondary antibodies (donkey anti-rabbit Alexa 488 and donkey anti-mouse Cy3) at 4 °C for 2 h
Sections were mounted in Vectashield anti-fading medium

controls

No positive or negative controls were explicitly mentioned in the input protocol.

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