Raf-1 Antibody | catalog# 50-237
Background
The Ras pathway is a critical signal transduction cascade involved in regulating cellular proliferation, differentiation, survival, and oncogenic transformation. Members of the Raf serine/threonine kinase family are key intermediates in this cascade, functioning to relay signals from activated Ras to the downstream protein kinases MEK and ERK (Marshall, 1996). Previous studies have shown that phosphorylation is required for Raf-1 activation (Dhillon and Kolch, 2002; Chong et al., 2003). Recent work has demonstrated that phosphorylation also regulates the downregulation of Raf (Dougherty et al., 2005) with two sites participating: Ser301 and Ser642.
Description
Western blot of UV treated human Jurkat cell lysate showing specific immunolabeling of the ~74k Raf-1 protein (Control). The phosphospecificity of this labeling is shown in the second lane (lambda-phosphatase: λ-Ptase). The blot is identical to the control except that it was incubated in λ-Ptase (1200 units for 30 min) before being exposed to the phospho Ser301 Raf-1 antibody. The immunolabeling of Raf-1 is completely eliminated by treatment with λ-Ptase.
Specifications
Clonality/Clone
This is a polyclonal antibody.Purification
Affinity PurifiedHost
Raf-1 antibody was raised in rabbit.Please use anti-rabbit secondary antibodies.
Immunogen
Phosphopeptide corresponding to amino acid residues surrounding the phospho-Ser301 of rat Raf-1.Application
The antibody has been directly tested for reactivity in Western blots with rat tissue. It is anticipated that the antibody will react with bovine, canine, chicken, human, mouse, non-human primate and Xenopus based on the fact that these species have 100% homology with the amino acid sequence used as antigen.Tested Application
WBBuffer
Raf-1 antibody is supplied as affinity purified polyclonal antibody, 100 μl in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 μg per ml BSA and 50% glycerol.Storage
Raf-1 antibody can be stored at -20˚C and is stable at -20˚C for at least 1 year.Positive Control
- Cat. No. 1205 - Jurkat Cell Lysate]
Species Reactivity
H, M, R, D, B, C, XReference
- Chong H, Vikis HG, Guan KL (2003) Mechanisms of regulating the Raf kinase family. Cellular Signalling 15:463-469.
- Dhillon AS, Kolch W (2002) Untying the regulation of the Raf-1 kinase. Arch Biochem Biophys 404:3-9.
- Dougherty MK, Muller J, Ritt DA, Zhou M, Zhou XZ, Copeland TD, Conrads TP, Veenstra TD, Lu KP, Morrison DK (2005) Regulation of Raf-1 by Direct Feedback Phosphorylation. Mol Cell 17:215-224.]