Cell Signaling Technology
Phospho-c-Jun (Ser63) (54B3) Rabbit mAb
2361
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c-Jun phosphorylated at Serine 63 by JNK. So the levels of Phospho-c-Jun (Ser63) is also used as a measure of activation status of JNK. When cells were treated with SP600125, the levels of Phospho-c-Jun (Ser63) were lower compared to DMSO treatment indicating JNK pathway inhibition.
Western Blot
CFBE41o- cells
Primary antibody (1:1000) in dilution buffer (TBST with 5% BSA) with gentle agitation overnight at 4°C.
TBST with 5% w/v nonfat dry milk
Incubated the membrane with the Goat Anti-Mouse IgG, H & L Chain Specific Peroxidase Conjugate (Cat No: 401253 Merckmillipore) 1:10000 diluted in 10 ml of blocking buffer with gentle agitation for 1 hr at room temperature.
None
Incubated the membrane with appropriate amount of Luminata Crescendo Western HRP substrate (WBLUR0100 Merckmillipore) for 5 min at room temperature. Drained the membrane of excess developing solution (without letting it to dry) wrapped in transparent plastic sheet and exposed the x-ray film for 1 min.
The antibody detects Phospho-C-JUN (ser63) in the blotting and its levels were lower upon treatment of SP600125 indicating the inactivation of JNK downstream pathway.
This antibody is also used in the published article PMID: 26701908
It works well as described
A good reagent to assay the Phospho-C-JUN by western blotting.