Protein Analysis via Immunoblotting and IP

Immunoblotting and immunoprecipitation were performed as previously described^{61 (link)}. The antibodies used in this study were: anti-dual phospho-JNK (Cell Signaling), anti-Flag (Sigma), anti-HA (Boehringer Mannheim), anti-phospho-Thr (Upstate), anti-phospho-Tyr (Upstate), anti-phospho-Ser (Upstate), anti-His (Sigma), anti-GST (GE), anti-JIP1 (Santa Cruz), and anti-myc (Upstate). In all cases, before performing gel-electrophoresis, the amount of total proteins was quantified using Bradford assay and equal amounts of proteins were loaded in each lane in a gel. All immunoblot analyses were independently performed at least three times.

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Moon J., Ha J, & Park S.H. (2017). Identification of PTPN1 as a novel negative regulator of the JNK MAPK pathway using a synthetic screening for pathway-specific phosphatases. Scientific Reports, 7, 12974.

Publication 2017

anti-Flag anti-HA anti-GST

Antibodies Assay Electrophoresis Immunoblot analyses Immunoprecipitation Proteins

Corresponding Organization :

Other organizations : Seoul National University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Phosphorylation of JNK (dual phospho-JNK)
Phosphorylation of proteins (phospho-Thr, phospho-Tyr, phospho-Ser)

control variables

Total protein amount quantified using Bradford assay and equal amounts of proteins loaded in each lane

controls

Positive controls: None specified
Negative controls: None specified

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