C-Raf (phospho-Ser-471) Blocking Peptide
€155.00
In stock
SKU
ECM-RX2905
Background:
The Ras-Raf-MAP kinase signaling pathway is involved in control of cell proliferation and differentiation. The Raf kinase family includes A-Raf, B-Raf, and C-Raf. Each family member has three highly conserved regions (CR1-3). The N-terminal CR1 contains the Ras-GTP-binding domain. The CR2 contains a negative regulatory serine residue (C-Raf (S259)/B-Raf(S365)) that may bind 14-3-3 proteins. The CR3 is the catalytic domain that contains phosphorylation sites for Raf-regulating enzymes within two segments, the N-region and the activation segment. Activation of C-Raf involves phosphorylation at many sites including Ser-338, Tyr-341, and Ser-471. The latter site is phosphorylated after EGF stimulation and may be important for MEK interaction in both C-Raf and A-Raf. In B-Raf, multiple phosphorylation sites have been identified, but their specific roles are uncertain. Phosphorylation of Ser-446 may prime B-Raf for activation, and Ser-446 and/or Ser-447 phosphorylation may be critical for B-Raf biological activity during PC12 differentiation. Ser-579 is required for growth factor activation and kinase activity.
Background References
Mason, C.S. et al. (1999) EMBOJ 18(8):2137.
Wilhelm, S.M. et al. (2004) Cancer Res 64:7099.
Zhu, J. et al. (2005) Mol. Biol. Cell 16:4733.
Brummer, T. et al. (2006) Oncogene 25(47):6262.
Karbowniczek, M. et al. (2006) J Biol Chem 281(35):25447.
Baljuls, A. et al. (2008) J Biol Chem 283(40):27239.
Sequence: A synthetic peptide corresponding to amino acid residues surrounding phosphorylated Ser-471 in human C-Raf. This sequence has high homology with similar regions in rat and mouse C-Raf, and has high homology to the conserved sites in B-Raf (Ser-579) and A-Raf (Ser-432).
Specificity: The peptide is specifically recognized by anti-C-Raf (Ser-471) phospho-specific antibody (RP2901) in ELISA, and has been shown to block the reactivity of RP2901 during Western blot. In addition, the peptide is recommended for use in blocking RP2901 reactivity in immunocytochemistry.
Application dilution:
Blocking: 1:1000
ELISA: 50 ng/well
End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1 hour at room temperature.
Buffer/Storage:
Blocking Peptide is supplied in 50µl phosphate-buffered saline and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
The Ras-Raf-MAP kinase signaling pathway is involved in control of cell proliferation and differentiation. The Raf kinase family includes A-Raf, B-Raf, and C-Raf. Each family member has three highly conserved regions (CR1-3). The N-terminal CR1 contains the Ras-GTP-binding domain. The CR2 contains a negative regulatory serine residue (C-Raf (S259)/B-Raf(S365)) that may bind 14-3-3 proteins. The CR3 is the catalytic domain that contains phosphorylation sites for Raf-regulating enzymes within two segments, the N-region and the activation segment. Activation of C-Raf involves phosphorylation at many sites including Ser-338, Tyr-341, and Ser-471. The latter site is phosphorylated after EGF stimulation and may be important for MEK interaction in both C-Raf and A-Raf. In B-Raf, multiple phosphorylation sites have been identified, but their specific roles are uncertain. Phosphorylation of Ser-446 may prime B-Raf for activation, and Ser-446 and/or Ser-447 phosphorylation may be critical for B-Raf biological activity during PC12 differentiation. Ser-579 is required for growth factor activation and kinase activity.
Background References
Mason, C.S. et al. (1999) EMBOJ 18(8):2137.
Wilhelm, S.M. et al. (2004) Cancer Res 64:7099.
Zhu, J. et al. (2005) Mol. Biol. Cell 16:4733.
Brummer, T. et al. (2006) Oncogene 25(47):6262.
Karbowniczek, M. et al. (2006) J Biol Chem 281(35):25447.
Baljuls, A. et al. (2008) J Biol Chem 283(40):27239.
Sequence: A synthetic peptide corresponding to amino acid residues surrounding phosphorylated Ser-471 in human C-Raf. This sequence has high homology with similar regions in rat and mouse C-Raf, and has high homology to the conserved sites in B-Raf (Ser-579) and A-Raf (Ser-432).
Specificity: The peptide is specifically recognized by anti-C-Raf (Ser-471) phospho-specific antibody (RP2901) in ELISA, and has been shown to block the reactivity of RP2901 during Western blot. In addition, the peptide is recommended for use in blocking RP2901 reactivity in immunocytochemistry.
Application dilution:
Blocking: 1:1000
ELISA: 50 ng/well
End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1 hour at room temperature.
Buffer/Storage:
Blocking Peptide is supplied in 50µl phosphate-buffered saline and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
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