Protocol detail

Quantifying Protein Expression via Western Blot

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Western blot analysis was performed as previously described [1 (link)] using BDNF (1:2,000; GeneTex, Irvine, CA, USA) and GAPDH (1:5000; Cell Signaling, Danvers, MA, USA) antibodies. Signal detection was performed using the LAS-2000 system (LAS-2000; Fuji, Tokyo, Japan) with enhanced chemiluminescence (Western-CDP star; PerkinElmer Life Sciences, Norwalk, CT, USA). Densitometric analysis of immunoreactivity for each protein was conducted using NIH ImageJ software.

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Lee B., Pothula S., Wu M., Kang H., Girgenti M.J., Picciotto M.R., DiLeone R.J., Taylor J.R, & Duman R.S. (2022). Positive modulation of N-methyl-D-aspartate receptors in the mPFC reduces the spontaneous recovery of fear. Molecular Psychiatry, 27(5), 2580-2589.

Publication 2022

BDNF GAPDH LAS-2000 system Western-CDP star

Antibodies Chemiluminescence Densitometric Gapdh Protein Signal detection Western blot analysis

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Variable analysis

independent variables

BDNF antibody (1:2,000; GeneTex, Irvine, CA, USA)

dependent variables

Densitometric analysis of immunoreactivity for each protein

control variables

GAPDH antibody (1:5000; Cell Signaling, Danvers, MA, USA)
Signal detection using the LAS-2000 system (LAS-2000; Fuji, Tokyo, Japan) with enhanced chemiluminescence (Western-CDP star; PerkinElmer Life Sciences, Norwalk, CT, USA)
Densitometric analysis using NIH ImageJ software

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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