Protein Isolation and Detection Workflow

Proteins were isolated and detected as described previously23 (link)24 (link)62 (link). Primary antibodies raised against following proteins were used: phospho-p38 MAPK (9212), phospho-MAPKAP kinase 2 (MK2) (3044), MAPKAP kinase 2 (3042), phospho γH2AX (9718), phospho-ATF2 (9221) from Cell Signaling Technology, Boston, MA), p38MAPKα (sc-535), phospho ERK (sc-16982R), ERK1 (sc-94), JNK (sc-571), ATF2 (sc-187) from Santa Cruz Biotechnology, Santa Cruz, CA), pJNK (AF1205), R&D Systems Minneapolis, MN, USA, GAPDH (AM4300, Ambion, Grand Island, NY), a-tubulin (T5168, Sigma Aldrich, Dorset, UK), Shc1 (610879), BD Biosciences, San Diego, CA, USA, pSer36-Shc1 (54518, Abcam, Cambridge, UK) and anti-HA-Peroxidase (12013819001), Roche, Mannheim, Germany. Antibodies were visualized by ECL Western blot detection reagents (Amersham, Buckinghamshire, UK), quantified by densitometric scanning using the image J program (NIH, Bethesda, MD) and normalized against loading control.

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Khalid S., Drasche A., Thurner M., Hermann M., Ashraf M.I., Fresser F., Baier G., Kremser L., Lindner H, & Troppmair J. (2016). cJun N-terminal kinase (JNK) phosphorylation of serine 36 is critical for p66Shc activation. Scientific Reports, 6, 20930.

Publication 2016

GAPDH (AM4300, pSer36-Shc1 anti-HA-Peroxidase

Antibodies Atf2 Erk1 Gapdh Mapkap kinase 2 P38 mapk Peroxidase Proteins Shc1 Tubulin Western blot

Corresponding Organization :

Other organizations : Innsbruck Medical University, Universität Innsbruck

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Variable analysis

independent variables

Proteins isolated and detected as described previously in the references

dependent variables

Levels of phospho-p38 MAPK, phospho-MAPKAP kinase 2 (MK2), MAPKAP kinase 2, phospho γH2AX, phospho-ATF2, p38MAPKα, phospho ERK, ERK1, JNK, ATF2, pJNK, GAPDH, α-tubulin, Shc1, pSer36-Shc1, and HA-Peroxidase

control variables

Loading control used for normalization of dependent variable measurements

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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