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Anti-Raf-1, clone AM223
Species Reactivity Key Applications Host Format Antibody Type
H, M, R IP, WB Rabbit null Monoclonal Antibody
Description:
Anti-Raf-1 Antibody, clone AM223 | 04-739
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Specificity:
Recognizes Raf-1, Mr 74 kDa.
Molecular Weight:
74 kDa
Epitope:
C-terminus
Immunogen:
KLH-conjugated, synthetic peptide corresponding to the C-terminus (amino acids 637-648 (CTLTTSPRLPVF)) of human Raf-1.
Clone:
AM223
Isotype:
IgG
Background Information:
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.
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Species Reactivity:
Human
Mouse
Rat
Species Reactivity Note:
Human and mouse. Predicted to cross-react with rat based on sequence homology.
Application Notes:
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.
Control:
3T3/A31 cell lysate.
Included Positive Antigen Control: Catalog # 12-305, 3T3/A31 lysate. Add 2.5 μL of 2-mercaptoethanol/100 μL of lysate and boil for 5 minutes to reduce the preparation. Load 20 μg of reduced lysate per lane for minigels.
Quality Assurance:
Routinely evaluated by Western Blot on 3T3/A31 lysates.
Presentation:
Cultured supernatant with 0.05% sodium azide.
Storage Conditions:
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.
UniProt Number:
P04049
Entrez Gene Number:
NM_002880
Gene Symbol:
C-RAF
CRAF
EC 2.7.11.1
NS5
RAF
Raf-1
c-Raf
cRaf
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Alternate Names:
Oncogene RAF1
raf proto-oncogene serine/threonine protein kinase
v-raf-1 murine leukemia viral oncogene homolog 1
Usage Statement:
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.
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Key Applications:
Immunoprecipitation
Western Blotting
Entrez Gene Summary:
This gene is the cellular homolog of viral raf gene (v-raf). The encoded protein is a MAP kinase kinase kinase (MAP3K), which functions downstream of the Ras family of membrane associated GTPases to which it binds directly. Once activated, the cellular RAF1 protein can phosphorylate to activate the dual specificitypro |