Product Pathways - MAPK Signaling
Phospho-c-Jun (Ser63) II Blocking Peptide #1020
Description
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.
Quality Control
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.
Western Blotting
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).
IHC-P (paraffin)
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).
Applications
Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry and Western blot protocols.
Directions for Use
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.
Background
c-Jun is a component of the transcription factor AP-1, which binds and activates transcription at TRE/AP-1 elements. The transcriptional activity of c-Jun is regulated by phosphorylation at Ser63 and Ser73 (1,2). Extracellular signals including growth factors, transforming oncoproteins and UV irradiation stimulate phosphorylation of c-Jun at Ser63/73 and activate c-Jun-dependent transcription. Mutation of Ser63/73 renders c-Jun nonresponsive to mitogenic and stress-induced signaling pathways. The MAP kinase homologue SAPK/JNK binds to the amino-terminal region of c-Jun and phosphorylates c-Jun at Ser63/73. In addition, the activity of SAPK/JNK is stimulated by the same signals that activate c-Jun (3,4).
- Binetruy, B. et al. (1991) Nature 351, 122-127.
- Smeal, T. et al. (1991) Nature 354, 494-496.
- Derijard, B. et al. (1994) Cell 76, 1025-1037.
- Kyriakis, J.M. et al. (1994) Nature 369, 156-160.
Application References
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