raf proto-oncogene serine/threonine protein kinase
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.
Product Information
Format
Purified
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.
Presentation
Cultured supernatant with 0.05% sodium azide.
Applications
Application
Anti-Raf-1 Antibody, clone AM223 is an antibody against Raf-1 for use in IP & WB.
Key Applications
Western Blotting
Immunoprecipitation
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.
Biological Information
Immunogen
KLH-conjugated, synthetic peptide corresponding to the C-terminus (amino acids 637-648 (CTLTTSPRLPVF)) of human Raf-1.
Epitope
C-terminus
Clone
AM223
Host
Rabbit
Specificity
Recognizes Raf-1, Mr 74 kDa.
Isotype
IgG
Species Reactivity
Rat Human Mouse
Species Reactivity Note
Human and mouse. Predicted to cross-react with rat based on sequence homology.
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 specificityprotein kinases MEK1 and MEK2, which in turn phosphorylate to activate the serine/threonine specific protein kinases, ERK1 and ERK2. Activated ERKs are pleiotropic effectors of cell physiology and play an important role in the control of gene expression involved in the cell division cycle, apoptosis, cell differentiation and cell migration. Mutations in this gene are associated with Noonan syndrome 5 and LEOPARD syndrome 2. [provided by RefSeq].
FUNCTION:Involved in the transduction of mitogenic signals from the cell membrane to the nucleus. Part of the Ras-dependent signaling pathway from receptors to the nucleus. Protects cells from apoptosis mediated by STK3. Ref.12
Catalytic activity ATP + a protein = ADP + a phosphoprotein.
Cofactor Binds 2 zinc ions per subunit.
SUBUNIT: Interacts with Ras proteins. The interaction with Ras is antagonized by RIN1. Weakly interacts with RIT1 By similarity. Interacts with STK3, which inhibits its pro-apoptotic activity.
SPECIFICITY:In skeletal muscle, isoform 1 is more abundant than isoform 2. Ref.7
PTM:Phosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylation at Thr-269 increases its kinase activity. Ref.6 Ref.8 Ref.9 Ref.13 Ref.14 Ref.15 Ref.16 Ref.17
DISEASE: Defects in RAF1 are the cause of Noonan syndrome type 5 (NS5) [MIM:611553]. Noonan syndrome (NS) is a disorder characterized by dysmorphic facial features, short stature, hypertelorism, cardiac anomalies, deafness, motor delay, and a bleeding diathesis. It is a genetically heterogeneous and relatively common syndrome, with an estimated incidence of 1 in 1000-2500 live births. Ref.22 Ref.23
Defects in RAF1 are the cause of LEOPARD syndrome type 2 (LEOPARD syndrome-2) [MIM:611554]. LEOPARD syndrome is an autosomal dominant disorder allelic with Noonan syndrome. The acronym LEOPARD stands for lentigines, electrocardiographic conduction abnormalities, ocular hypertelorism, pulmonic stenosis, abnormalities of genitalia, retardation of growth, and deafness.
SIMILARITY:Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. RAF subfamily.
Contains 1 phorbol-ester/DAG-type zinc finger.
Contains 1 protein kinase domain.
Contains 1 RBD (Ras-binding) domain.
Product Usage Statements
Quality Assurance
Routinely evaluated by Western Blot on 3T3/A31 lysates.
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.
Storage and Shipping Information
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.
raf proto-oncogene serine/threonine protein kinase
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.
Product Information
Format
Purified
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.
Presentation
Cultured supernatant with 0.05% sodium azide.
Applications
Application
Anti-Raf-1 Antibody, clone AM223 is an antibody against Raf-1 for use in IP & WB.
Key Applications
Western Blotting
Immunoprecipitation
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.
Biological Information
Immunogen
KLH-conjugated, synthetic peptide corresponding to the C-terminus (amino acids 637-648 (CTLTTSPRLPVF)) of human Raf-1.
Epitope
C-terminus
Clone
AM223
Host
Rabbit
Specificity
Recognizes Raf-1, Mr 74 kDa.
Isotype
IgG
Species Reactivity
Rat Human Mouse
Species Reactivity Note
Human and mouse. Predicted to cross-react with rat based on sequence homology.
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 specificityprotein kinases MEK1 and MEK2, which in turn phosphorylate to activate the serine/threonine specific protein kinases, ERK1 and ERK2. Activated ERKs are pleiotropic effectors of cell physiology and play an important role in the control of gene expression involved in the cell division cycle, apoptosis, cell differentiation and cell migration. Mutations in this gene are associated with Noonan syndrome 5 and LEOPARD syndrome 2. [provided by RefSeq].
FUNCTION:Involved in the transduction of mitogenic signals from the cell membrane to the nucleus. Part of the Ras-dependent signaling pathway from receptors to the nucleus. Protects cells from apoptosis mediated by STK3. Ref.12
Catalytic activity ATP + a protein = ADP + a phosphoprotein.
Cofactor Binds 2 zinc ions per subunit.
SUBUNIT: Interacts with Ras proteins. The interaction with Ras is antagonized by RIN1. Weakly interacts with RIT1 By similarity. Interacts with STK3, which inhibits its pro-apoptotic activity.
SPECIFICITY:In skeletal muscle, isoform 1 is more abundant than isoform 2. Ref.7
PTM:Phosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylation at Thr-269 increases its kinase activity. Ref.6 Ref.8 Ref.9 Ref.13 Ref.14 Ref.15 Ref.16 Ref.17
DISEASE: Defects in RAF1 are the cause of Noonan syndrome type 5 (NS5) [MIM:611553]. Noonan syndrome (NS) is a disorder characterized by dysmorphic facial features, short stature, hypertelorism, cardiac anomalies, deafness, motor delay, and a bleeding diathesis. It is a genetically heterogeneous and relatively common syndrome, with an estimated incidence of 1 in 1000-2500 live births. Ref.22 Ref.23
Defects in RAF1 are the cause of LEOPARD syndrome type 2 (LEOPARD syndrome-2) [MIM:611554]. LEOPARD syndrome is an autosomal dominant disorder allelic with Noonan syndrome. The acronym LEOPARD stands for lentigines, electrocardiographic conduction abnormalities, ocular hypertelorism, pulmonic stenosis, abnormalities of genitalia, retardation of growth, and deafness.
SIMILARITY:Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. RAF subfamily.
Contains 1 phorbol-ester/DAG-type zinc finger.
Contains 1 protein kinase domain.
Contains 1 RBD (Ras-binding) domain.
Product Usage Statements
Quality Assurance
Routinely evaluated by Western Blot on 3T3/A31 lysates.
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.
Storage and Shipping Information
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.
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