Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-c-Jun (Ser63) (54B3) Rabbit mAb in the presence of control peptide (left) or Phospho-c-Jun (Ser63) II Blocking Peptide (#1020) (right).
Western blot analysis of whole cell lysates from NIH/3T3 cells treated and untreated with UV light, probed with Phospho-c-Jun (Ser63) II Antibody #9261 (left) and with the same antibody preincubated with antigen-specific blocking peptide (right).
For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 ul total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the relevant product data sheet.
For Western immunoblotting, add 10 µl of antibody and 10 µl of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room temperature for 30 minutes before allowing to react with the blot.
Supplied in 20 mM potassium phosphate (pH 7.0), 50 mM NaCl, 0.1 mM EDTA, 1 mg/ml BSA and 5% glycerol. Store at –20°C.
This peptide is used to block Phospho-c-Jun (Ser63) (54B3) Rabbit mAb #2361 reactivity. This peptide will also block Phospho-c-Jun (Ser63) II Antibody #9261 reactivity by Western.
The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Phospho-c-Jun (Ser63) (54B3) Rabbit mAb #2361 by immunohistochemistry and Western blotting. The peptide also blocks Phospho-c-Jun (Ser63) II Antibody #9261 by Western blotting.
c-Jun is a member of the Jun family containing c-Jun, JunB, and JunD, and is a component of the transcription factor activator protein-1 (AP-1). AP-1 is composed of dimers of Fos, Jun, and ATF family members and binds to and activates transcription at TRE/AP-1 elements (reviewed in 1). Extracellular signals including growth factors, chemokines, and stress activate AP-1-dependent transcription. The transcriptional activity of c-Jun is regulated by phosphorylation at Ser63 and Ser73 through SAPK/JNK (reviewed in 2). Knock-out studies in mice have shown that c-Jun is essential for embryogenesis (3), and subsequent studies have demonstrated roles for c-Jun in various tissues and developmental processes including axon regeneration (4), liver regeneration (5), and T cell development (6). AP-1 regulated genes exert diverse biological functions including cell proliferation, differentiation, and apoptosis, as well as transformation, invasion and metastasis, depending on cell type and context (7-9). Other target genes regulate survival, as well as hypoxia and angiogenesis (8,10). Research studies have implicated c-Jun as a promising therapeutic target for cancer, vascular remodeling, acute inflammation, and rheumatoid arthritis (11,12).
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