General description
The Raf proteins (Raf-1, A-Raf, B-Raf) are Ser/Thr kinases with homology to the PKC family, containing an N-terminal regulatory domain and a c-terminal catalytic domain. Members of the Raf family bind to activated Ras GTPase, which results in Raf translocation to the plasma membrane and activation. Activated Raf proteins phosphorylate MEKs, and are therefore the principle transducers of signals from Ras to MAP kinase. In addition to its role in mitogenesis, Raf-1 may play a role in regulation of apoptosis and cell cycle progression. Activation of Raf-1 involves phosphorylation of Ser338/339 and Tyr340/341. Activating mutations of B-Raf that disrupt its auto-inhibition loop have been implicated in a number of cancers, including melanoma and colon cancer.
Specificity
Predicted to cross-react with rat based on sequence homology.
Recognizes Raf-1, Mr 74 kDa.
Immunogen
Epitope: C-terminus
KLH-conjugated, synthetic peptide corresponding to the C-terminus (amino acids 637-648 (CTLTTSPRLPVF)) of human Raf-1.
Application
Anti-Raf-1 Antibody, clone AM223 is an antibody against Raf-1 for use in IP & WB.
Research Category
Signaling
Research Sub Category
MAP Kinases
Western Blotting Analysis:
A 1:500-1:2,000 diltuoin of this lot detected Raf-1 in 3T3/A31 cell lysates.
Immunoprecipitation: 2-4 μg of a previous lot immunoprecipitated Raf-1 from 400 μg of 3T3/A31 RIPA lysate.
Quality
Routinely evaluated by Western Blot on 3T3/A31 lysates.
Target description
74 kDa
Linkage
Replaces: 04-412
Physical form
Cultured supernatant with 0.05% sodium azide.
Storage and Stability
Stable for 1 year at -20ºC from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Analysis Note
Control
Positive Antigen Control: Catalog #12-305, 3T3/A31 lysate. Add 2.5 μL of 2-mercapto-ethanol/100 μL of lysate and boil for 5 minutes to reduce the preparation. Load 20 μg of reduced lysate per lane for minigels.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
- UPC:
- 51202301
- Condition:
- New
- Weight:
- 1.00 Ounces
- HazmatClass:
- No
- WeightUOM:
- LB
- MPN:
- 04-739
- Product Size:
- 739