Protocol detail

Western Blot Analysis of Molecular Targets

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Whole protein extracts and Western blot (WB) analysis were performed using standard procedures as described in our previous study,^{9 (link)} with detection using the enhanced chemiluminescence system (Life Technologies, Grand Island, NY). Antibodies against SLC1A5 were used at 1:1,000 dilutions (Millipore, Billerica, MA), SN1 at 1:100 (Epitomics, Burlin-game, CA), LAT1 at 1:1,000 (Cell Signaling Technology, Danvers, MA), Caspase 3 at 1:1,000 (Cell Signaling Technology), Caspase 9 at 1:1,000 (Cell Signaling Technology), LC3 1:1,000 (Medical and Biological Laboratories, Nagoya, Japan), Bcl-2 1:1,000 (Cell Signaling Technology) and Bax 1:1000 (Cell Signaling Technology).

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Hassanein M., Qian J., Hoeksema M.D., Wang J., Jacobovitz M., Ji X., Harris F.T., Harris B.K., Boyd K.L., Chen H., Eisenberg R, & Massion P.P. (2015). Targeting SLC1A5-mediated glutamine dependence in non-small cell lung cancer. International journal of cancer. Journal international du cancer, 137(7), 1587-1597.

Publication 2015

enhanced chemiluminescence system Caspase 3 Caspase 9 Bcl-2 Bax

Antibodies Bcl 2 Biological Caspase 1 Chemiluminescence Dilutions Protein Western blot analysis

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Variable analysis

independent variables

Protein expression levels of SLC1A5, SN1, LAT1, Caspase 3, Caspase 9, LC3, Bcl-2, and Bax

dependent variables

Not explicitly mentioned

control variables

Experimental procedures were performed using standard procedures as described in a previous study
Detection was done using the enhanced chemiluminescence system

controls

Positive control: Not specified
Negative control: Not specified

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