Immunohistochemistry for NeuN and ID1

Immunohistochemistry for NeuN (a neuronal marker) and ID1 was performed using a published procedure (28 (link)). The sections were incubated in mouse anti-NeuN (1:800; MAB377; EMD Millipore, Billerica, MA, USA) or rabbit anti-ID1 (1:200; sc-488; Santa Cruz Biotechnology, Inc.) as primary antibodies, and in biotinylated horse anti-mouse (1:200; BA-2001; Vector Laboratories, Inc., Burlingame, CA, USA or rabbit anti-goat immunoglobulin G (1:200; BA-1100; Vector Laboratories, Inc.) and streptavidin peroxidase complex (VECTASTAIN^® Elite ABC kit 1:200; Vector Laboratories, Inc.) as secondary antibodies. The antibodies were finally visualized with 3,3′-diaminobenzidine tetrahydrochloride. A negative control test was performed to establish the specificity of the immunostaining using pre-blocking serum (S-1000; Vector Laboratories, Inc.) instead of primary antibody. The negative control test showed no immunoreactivity in structures observed.

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Lee J.C., Kim I.H., Cho J.H., Lee T.K., Park J.H., Ahn J.H., Shin B.N., Yan B.C., Kim J.D., Jeon Y.H., Lee Y.J., Won M.H, & Kang I.J. (2018). Vanillin improves scopolamine-induced memory impairment through restoration of ID1 expression in the mouse hippocampus. Molecular Medicine Reports, 17(3), 4399-4405.

Publication 2018

MAB377 sc-488 BA-2001 BA-1100 VECTASTAIN® Elite ABC kit S-1000

Abc 1 Anti immunoglobulin Antibodies Antibody Goat Horse Immunohistochemistry Mouse Neuronal Peroxidase Rabbit Serum Streptavidin Vector

Corresponding Organization :

Other organizations : Kangwon National University, Hallym University, Yangzhou University

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Variable analysis

independent variables

Antibody type (mouse anti-NeuN, rabbit anti-ID1)

dependent variables

Immunoreactivity (presence/absence) in tissue sections

control variables

Antibody concentration (1:800 for NeuN, 1:200 for ID1)
Secondary antibody type (biotinylated horse anti-mouse, rabbit anti-goat IgG)
Secondary antibody concentration (1:200)
Visualization method (streptavidin peroxidase complex, 3,3′-diaminobenzidine tetrahydrochloride)

controls

Positive control: Tissue sections incubated with primary antibodies (mouse anti-NeuN, rabbit anti-ID1)
Negative control: Tissue sections incubated with pre-blocking serum instead of primary antibody

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