Cells were collected in sodium dodecyl sulfate sample buffer. After sonication, cell lysates were subjected to electrophoresis on 5–20% polyacrylamide gels and transferred onto polyvinylidene difluoride membranes (ATTO). The membranes were probed with specific antibodies for poly(adenosine diphosphate-ribose) polymerase (PARP), JNK, c-Jun, phosphorylated-c-Jun (Ser63) (Cell Signaling Technology, Danvers, MA, USA) and GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA). After washing, the membranes were incubated with secondary horse-radish peroxidase-conjugated antibodies. Signals were detected by means of enhanced chemiluminescence (GE Healthcare Japan, Tokyo, Japan) and scanned with an image analyzer LAS-4000 and Image Gauge (version 3.1) (Fuji Film, Tokyo, Japan). Band intensities were determined using ImageJ software [14 (link)].
Phosphorylated c jun ser63
Manufactured by Cell Signaling Technology
Sourced in United States
Phosphorylated-c-Jun (Ser63) is a laboratory reagent used to detect and quantify the phosphorylation of the c-Jun protein at serine 63. This phosphorylation event is a key regulatory mechanism in cellular signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Las 4000, by Fujifilm (1 mentions)
Enhanced chemiluminescence, by GE Healthcare (1 mentions)
Image gauge version 3.1, by Fujifilm (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Erk1 2, by Cell Signaling Technology (1 mentions)
Phospho erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
Bca kit, by Thermo Fisher Scientific (1 mentions)
Cyclin d1, by Abcam (1 mentions)
β actin, by Abcam (1 mentions)
C jun, by Cell Signaling Technology (1 mentions)
2 protocols using phosphorylated c jun ser63
1
Western Blot Analysis of Cellular Proteins
2
Protein Expression Analysis by Western Blot
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Total cellular protein was extracted using RIPA lysis containing protease and phosphatase inhibitors. Protein concentration was measured according to the instructions of the BCA kit (Thermo Scientific). SDS-PAGE separation and im-munoblotting were then performed according to standard methods. The primary antibodies used in our study are listed below: EK2 (Abcam), ERK1/2 (Cell Signaling Technology), c-JUN (Cell Signaling Technology), phospho-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology), phosphorylated c-JUN (Ser63) (Cell Signaling Technology), cyclin D1 (Abcam), and β-actin (Abcam).
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