R0495-06
Raf1, CT
Size
£495.00
- SKU:
- R0495-06
- Purity:
- Purified by Protein A affinity chromatography.
- Storage Conditions:
- -20°C
- Supplier:
- United States Biological
- Host:
- Sheep
- Immunogen:
- Synthetic peptide corresponding to aa637-648, CTLTTSPRLPVF of human Raf-1, conjugated to KLH. Species Sequence Homology: rat
- Format:
- Supplied as a solution in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, 30% glycerol
- Extra Details:
- Raf-1 associates with members of the 14–3–3 family of proteins, which appear to protect active Raf-1 from phosphatase action. Important regulatory phosphorylation events occur on tyrosines 340 and 341 and serines 259, 499 (activating), serine 43 (prevents Ras:GTP binding), and serine 621 (constitutive, required for activity). Once active, Raf-1 associates with plasma membranes independent of Ras and phosphorylates and activates MEK, initiating the MAP kinase signaling pathway. Applications: Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested. Recommended Dilutions: Western Blot: 0.5-2ug/ml detects Raf-1 in RIPA lysates from 3T3/A31 cell lysates. 3T3/A31 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with R0495-06 (2ug/ml). Proteins were visualized using an IgG (HRP) goat anti-rabbit secondary antibody and a chemiluminescence detection system. Immunoprecipitation: 4ug immunoprecipitated Raf-1. The immunocomplex was then used in a coupled phosphorylation assay to activate MEK1, which in turn activates MAP Kinase 2 that phosphorylates myelin basic protein in vitro. Optimal dilutions to be determined by the researcher. Included Positive Antigen Control: F4209: Fibroblast Cell, 3T3/A31. Add 2.5ul of 2-mercaptoethanol/100ul of lysate and boil for 5 minutes to reduce the preparation. Load 20ug of reduced lysate per lane for minigels. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
- Shipping Conditions:
- Blue Ice