Western Blotting Analysis of Protein Expression

Western blotting analysis was conducted with 20 μg of protein as described previously 14 (link)-16 (link). Primary antibodies were rabbit anti-TRAM2 antibody (dilute concentration, 1:1000, Thermo Fisher Scientific), mouse anti-GAPDH antibody (dilute concentration, 1:1000, Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-protein kinase RNA(PKR) - like ER kinase (PERK) antibody (dilute concentration, 1:1000, Santa Cruz Biotechnology), rabbit anti-MT1-MMP antibody (dilute concentration, 1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-MMP2 antibody (dilute concentration, 1:500, Merck KGaA, Darmstadt, Germany), and rabbit anti-MMP9 antibody (dilute concentration, 1:3000, GeneTex Inc., Irvine, CA, USA). Horseradish peroxidase-conjugated anti-rabbit or anti-mouse IgG (Promega, Madison, WI, USA) was used as secondary antibodies.

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Fukushima R., Kasamatsu A., Nakashima D., Higo M., Fushimi K., Kasama H., Endo-Sakamoto Y., Shiiba M., Tanzawa H, & Uzawa K. (2018). Overexpression of Translocation Associated Membrane Protein 2 Leading to Cancer-Associated Matrix Metalloproteinase Activation as a Putative Metastatic Factor for Human Oral Cancer. Journal of Cancer, 9(18), 3326-3333.

Publication 2018

mouse anti-GAPDH antibody Horseradish peroxidase-conjugated anti-rabbit or anti-mouse IgG

Anti antibody Anti igg Antibodies Antibody Dilute Gapdh Horseradish peroxidase Kinase Mmp2 Mmp9 Mouse Promega Protein Protein kinase Rabbit Western blotting analysis

Corresponding Organization :

Other organizations : Chiba University, Chiba University Hospital, Teikyo University Chiba Medical Center

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Variable analysis

independent variables

Amount of protein used for Western blotting analysis (20 μg)

dependent variables

Protein expression levels of TRAM2, GAPDH, PERK, MT1-MMP, MMP2, and MMP9 as measured by Western blotting

control variables

Experimental procedures as described previously in references 14-16

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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