Raf-1 phosphorylates and activates MEK-1, a kinase that activates the extracellular signal-regulated kinases (ERK). The Ras/Raf/MEK/ERK module is a ubiquitously expressed signaling pathway that conveys mitogenic and differentiation signals from the cell membrane to the nucleus1. This kinase cascade controls the proliferation and differentiation of different cell types. The small G protein Ras is activated by several growth factor receptors, and when activated binds Raf-1 kinase with high affinity. This induces the recruitment of Raf-1 from the cytosol to the cell membrane and its subsequent activation. Activated Raf-1 then phosphorylates the kinase MEK, thus activating it. MEK in turn phosphorylates and activates ERK, the prototypic mitogen-activated protein kinase (MAPK). Activated ERKs can translocate to the nucleus and regulate gene expression by the phosphorylation of transcription factors. Raf kinase inhibitor protein (RKIP) is a Raf-1 interacting protein that inhibits the phophorylation and activation of MEK by Raf-1. RKIP is not a substrate for either Raf-1 or MEK, but competitively disrupts the interaction between these kinases.

Suitable for use in Immunoprecipitation and Western Blot. Other applications have not been tested.

Immunoprecipitation: 7-10ug/IP reaction Western Blot: 1-3ug/ml Optimal dilutions to be determined by the researcher.

Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: MaxLight™650 conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

Note: Applications are based on unconjugated antibody.

Synthetic peptide derived from the C-terminal region of human RKIP protein.

Supplied as a liquid in PBS, pH 7.2. No preservatives added. Labeled with MaxLight™650.

Purifed by Protein A chromatography

Specific for the C-terminal region of the RKIP protein. On Western blots, it identifies a band at 30kDa in RKIP-transfected HEK293 lysates and 23kDa in HEK293 lysates or total mouse brain lysates. Reactivity has been confirmed with HEK 293 lysates. Immunoprecipitation tests with HEK293 cell preparations also yielded the ~23kD band.