Protein Expression Analysis in Hippocampus

Western blotting was conducted according to previously published methodology [8 (link),16 (link)]. The hippocampal tissues were homogenized on ice and lysed in a lysis buffer. Protein (30 μg) was separated on sodium dodecyl sulfate-polyacrylamide gels and transferred onto a nitrocellulose membrane. Mouse beta-actin antibody (1:1,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse GAP43 antibody (1:1,000; Santa Cruz Biotechnology), rabbit BDNF antibody (1:500; Santa Cruz Biotechnology), rabbit TrkB antibody (1:1,000; Santa Cruz Biotechnology), rabbit PSD95 antibody (1:1,000; Santa Cruz Biotechnology), rabbit Egr-1 antibody (1:1,000; Santa Cruz Biotechnology), and goat pCREB antibody (1:1,000; Santa Cruz Biotechnology) were used as the primary antibodies. Horseradish peroxidase-conjugated anti-rabbit antibody for BDNF, TrkB, PSD95, and Egr-1 (1:3,000; Vector Laboratories, Burlingame, CA, USA), horseradish peroxidase-conjugated anti-mouse antibody for beta-actin and GAP43 (1:2,000; Vector Laboratories), and horseradish peroxidase-conjugated anti-goat antibody for p-CREB (1:5,000; Santa Cruz Biotechnology) were used as the secondary antibodies. Band detection was conducted using an enhanced chemiluminescence detection kit (Santa Cruz Biotechnology).

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Hwang D.S., Kwak H.B., Ko I.G., Kim S.E., Jin J.J., Ji E.S., Choi H.H, & Kwon O.Y. (2016). Treadmill Exercise Improves Memory Function Depending on Circadian Rhythm Changes in Mice. International Neurourology Journal, 20(Suppl 2), S141-149.

Publication 2016

Mouse beta-actin antibody rabbit BDNF antibody rabbit TrkB antibody enhanced chemiluminescence detection kit

Anti antibody Antibodies Antibody Beta actin Buffer Chemiluminescence Egr 1 Goat Horseradish peroxidase Membrane Mouse Nitrocellulose Polyacrylamide gels Protein Rabbit Sodium dodecyl sulfate Tissues Trkb Vector

Corresponding Organization :

Other organizations : Kyung Hee University, Inha University, Dongseo University, Kyung Hee University Medical Center

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein levels of beta-actin, GAP43, BDNF, TrkB, PSD95, Egr-1, and pCREB

control variables

Protein load (30 μg) for western blotting
Primary and secondary antibodies used for western blotting

controls

Positive control: None mentioned
Negative control: None mentioned

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