Product Pathways - MAPK Signaling
Phospho-p38 MAPK (Thr180/Tyr182) Blocking Peptide #1170
Description
This peptide is used to specifically block Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb (IHC Preferred) #4631 reactivity.
Quality Control
The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb (IHC Preferred) #4631 signal completely in both immunohistochemistry and Western blotting. The peptide also blocks Phospho-p38 MAP Kinase (Thr180/Tyr182) (3D7) Rabbit mAb #9215 and Phospho-p38 MAP Kinase (Thr180/Tyr182) (28B10) Mouse mAb # 9216 signals in Western blotting.
Western Blotting
Western blot analysis of extracts from NIH/3T3 cells treated with UV, and C6 cells treated with anisomycin, using Phospho-p38 MAPK (Thr180/Tyr182) (3D7) Rabbit mAb #9215 (upper) or the same antibody preincubated with Phospho-p38 MAPK (Thr180/Tyr182) Blocking Peptide (lower).
Western Blotting
Western blot analysis of extracts from NIH/3T3 cells treated with UV, and C6 cells treated with anisomycin, using Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb (IHC preferred) #4631 (upper) or the same antibody preincubated with Phospho-p38 MAPK (Thr180/Tyr182) Blocking Peptide (lower).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb (IHC Preferred) #4631 preincubated with an irrelevant control peptide (left), or with Phospho-p38 MAPK (Thr180/Tyr182) Blocking Peptide (right).
Table
Phospho-p38 MAPK (Thr180/Tyr182) Blocking Peptide can be used to block signal from the indicated phospho-p38 MAPK antibodies in the indicated applications.
Applications
Use as a blocking reagent to evaluate the specificity of antibody reactivity in Western immunoblotting and immunohistochemistry protocols.
Directions for Use
For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 µl 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
p38 MAP kinase (MAPK), also called RK (1) or CSBP (2), is the mammalian orthologue of the yeast HOG kinase which participates in a signaling cascade controlling cellular responses to cytokines and stress (1-4). Four isoforms of p38 MAP kinase, p38α, β, γ (also known as ERK6 or SAPK3) and δ (also known as SAPK4) have been identified. Similar to the SAPK/JNK pathway, p38 MAP kinase is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharides (LPS), UV light and growth factors (1-5). MKK3, MKK6 and SEK activate p38 MAP kinase by phosphorylation at Thr180 and Tyr182. Activated p38 MAP kinase has been shown to phosphorylate and activate MAPKAP kinase 2 (3) and to phosphorylate the transcription factors ATF-2 (5), Max (6) and MEF2 (5-8).
- Rouse, J. et al. (1994) Cell 78, 1027-1037.
- Han, J. et al. (1994) Science 265, 808-811.
- Lee, J.C. et al. (1994) Nature 372, 739-746.
- Freshney, N.W. et al. (1994) Cell 78, 1039-1049.
- Raingeaud, J. et al. (1995) J. Biol. Chem. 270, 7420-7426.
- Zervos, A.S. et al. (1995) Proc. Natl. Acad. Sci. USA 92, 10531-10534.
- Zhao, M. et al. (1999) Mol. Cell. Biol. 19, 21-30.
- Yang, S.H. et al. (1999) Mol. Cell. Biol. 19, 4028-4038.
Application References
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Companion Products
- 4631 Phospho-p38 MAPK (Thr180/Tyr182)(12F8) Rabbit mAb (IHC Preferred)
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
