Western Blot Analysis of Protein Markers

Protein extracts were prepared as previously described^{22 (link)}. Overall, 10 μg of samples were electrophoresed and blotted to PVDF membrane (Bio-Rad, Hercules, CA, USA). Western blot was performed using primary antibodies against 15-lipoxygenase-1 (Mouse, Abcam, Cambridge, UK), PLIN1 (Rabbit, Cell Signaling, Danvers, MA, USA), Keratin 14 (Rabbit, Cell Signaling), phospho-RIP3(Rabbit, Abcam), RIP3(Rabbit, Cell signaling), ZBP1 (Rabbit, Cell Signaling), cleaved Caspase-3 (Asp175) (Rabbit, Cell Signaling), Caspase 3(Rabbit, Cell Signaling), Caspase 7(Rabbit, Cell Signaling), Tubulin (Cell Signaling), and β-actin (Mouse, Santa Cruz Biotechnology, Dallas, TX, USA) and secondary anti-mouse and anti-rabbit horse-radish peroxidase antibodies (Cell Signaling) as described previously. The blots were visualized with SuperSignal West Dura Substrate (ThermoFisher Scientific).

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Kim S.N., Akindehin S., Kwon H.J., Son Y.H., Saha A., Jung Y.S., Seong J.K., Lim K.M., Sung J.H., Maddipati K.R, & Lee Y.H. (2018). Anti-inflammatory role of 15-lipoxygenase contributes to the maintenance of skin integrity in mice. Scientific Reports, 8, 8856.

Publication 2018

PVDF membrane 15-lipoxygenase-1 PLIN1 Keratin 14 phospho-RIP3 cleaved Caspase-3 (Asp175) Caspase 3 Caspase 7 Tubulin β-actin secondary anti-mouse and anti-rabbit horse-radish peroxidase antibodies SuperSignal West Dura Substrate

15 lipoxygenase Actin Anti antibodies Antibodies Caspase 3 Caspase 7 Dura Horse radish peroxidase Keratin 14 Lipoxygenase 1 Membrane Mouse Protein Pvdf Rabbit Rip3 Tubulin Western blot

Corresponding Organization :

Other organizations : Yonsei University, Pusan National University, Seoul National University, Ewha Womans University, Wayne State University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein levels of 15-lipoxygenase-1, PLIN1, Keratin 14, phospho-RIP3, RIP3, ZBP1, cleaved Caspase-3, Caspase 3, Caspase 7, Tubulin, and β-actin

control variables

None explicitly mentioned

controls

Positive control: None mentioned
Negative control: None mentioned

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