Western blot of UV treated human Jurkat cell lysate showing specific immunolabeling of the ~74 kDa Raf-1 protein (Lane 1). The phosphospecificty of this labeling is shown in Lane 2 (lambda-phosphatase). The blot is identical to the control except that it was incubated in lambda-Ptase (1200 units for 30 min) before being exposed to the Anti-Ser301 Raf-1. The immunolabeling of Raf-1 is completely eliminated by treatment with lambda-Ptase.
|Tested species reactivity||Human, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic phospho-peptide corresponding to amino acid residues surrounding Ser301 of rat RAF1 conjugated to KLH|
|Storage buffer||0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/ml BSA, 50% glycerol|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody is predicted to react with bovine, canine, chicken, mouse, non-human primate and Xenopus based on 100% sequence homology.
This antibody is specific for the ~74 kDa Raf-1 protein phosphorylated at Ser301. Immunolabeling can be blocked by lambda-phosphatase treatment.
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; Chon et al, 2003). Recent work has demonstrated that phosphorylation also regulates the downregulation of Raf (Dougherty et al, 2005) with two sites participating Ser 301 and Ser 642.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.