Recombinant Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3433(2)] to Raf1 (phospho S259)
- Suitable for: WB, Dot blot
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Raf1 (phospho S259) antibody [EPR3433(2)]
See all Raf1 primary antibodies -
Description
Rabbit monoclonal [EPR3433(2)] to Raf1 (phospho S259) -
Host species
Rabbit -
Tested applications
Suitable for: WB, Dot blotmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Treated C6, NIH/3T3 and HEK-293 whole cell lysates.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3433(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab173539 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000 - 1/5000. Predicted molecular weight: 73 kDa.
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Dot blot |
Use at an assay dependent concentration.
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Notes |
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WB
1/1000 - 1/5000. Predicted molecular weight: 73 kDa. |
Dot blot
Use at an assay dependent concentration. |
Target
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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. -
Tissue specificity
In skeletal muscle, isoform 1 is more abundant than isoform 2. -
Involvement in 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.
Defects in RAF1 are the cause of LEOPARD syndrome type 2 (LEOPARD2) [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. -
Sequence similarities
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. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylation at Thr-269 increases its kinase activity. Phosphorylation at Ser-259 induces the interaction with YWHAZ and inactivates kinase activity. Dephosphorylation of Ser-259 by the complex containing protein phosphatase 1, SHOC2 and M-Ras/MRAS relieves inactivation, leading to stimulate RAF1 activity. -
Cellular localization
Cytoplasm. Cell membrane. Colocalizes with RGS14 and BRAF in both the cytoplasm and membranes. - Information by UniProt
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Database links
- Entrez Gene: 5894 Human
- Entrez Gene: 110157 Mouse
- Entrez Gene: 24703 Rat
- Omim: 164760 Human
- SwissProt: P04049 Human
- SwissProt: Q99N57 Mouse
- SwissProt: P11345 Rat
- Unigene: 159130 Human
see all -
Alternative names
- c Raf antibody
- C-raf antibody
- C-Raf proto-oncogene, serine/threonine kinase antibody
see all
Images
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All lanes : Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/5000 dilution (Purified)
Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) treated with 100nM Phorbol-12-myristate-13-acetate for 1 hour whole cell lysates
Lane 3 : HEK-293 (Human embryonic kidney epithelial cell) treated with 100nM Phorbol-12-myristate-13-acetate for 1 hour whole cell lysates, then incubated with phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 73 kDa
Observed band size: 73 kDaab181602 was used as GAPDH loading control.
ab181115 was used to detect Raf1.
ab150365 was used to detect Raf1 phospho S43.
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All lanes : Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/1000 dilution (Purified)
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 5uM forskolin for 10 minutes whole cell lysates
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) treated with 5uM forskolin for 10 minutes whole cell lysates, then incubated with phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 73 kDa
Observed band size: 73 kDa -
Lane 1 : Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/1000 dilution (Purified)
Lanes 2-3 : Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/1000 dilution
Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysates
Lane 2 : C6 (Rat glial tumor glial cell) treated with 30uM forskolin for 20 minutes whole cell lysates
Lane 3 : C6 (Rat glial tumor glial cell) treated with 30uM forskolin for 20 minutes whole cell lysates, then incubated with phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 73 kDa
Observed band size: 73 kDa -
All lanes : Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/5000 dilution
Lane 1 : HEK293 whole cell lysate
Lane 2 : HEK293 whole cell lysate treated with 100nM Phorbol-12-myristate-13-acetate for 1 hour
Lane 3 : HEK293 whole cell lysate treated with 100nM Phorbol-12-myristate-13-acetate for 1 hour then incubated with phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 73 kDa
Observed band size: 73 kDa
Exposure time: 1 secondBlocking and dilution buffer: 5% NFDM/TBST.
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Dot blot analysis of Raf1 (pS259) peptide (Lane 1) and Raf1 non-phospho peptide (Lane 2) labelling Raf1 (pS259) with ab173539 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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All lanes : Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/1000 dilution
Lane 1 : 293T cell lysates
Lane 2 : 293T cell lysates treated with Okadaic Acid + Calyculin A
Lysates/proteins at 10 µg per lane.
Predicted band size: 73 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (13)
ab173539 has been referenced in 13 publications.
- Li W et al. 4,5-Dimethoxycanthin-6-one is a novel LSD1 inhibitor that inhibits proliferation of glioblastoma cells and induces apoptosis and pyroptosis. Cancer Cell Int 22:32 (2022). PubMed: 35042538
- Li J et al. METTL3 Facilitates Tumor Progression by COL12A1/MAPK Signaling Pathway in Esophageal Squamous Cell Carcinoma. J Cancer 13:1972-1984 (2022). PubMed: 35399719
- Bai D et al. BPTF activates the MAPK pathway through coexpression with Raf1 to promote proliferation of T-cell lymphoma. Oncol Lett 24:223 (2022). PubMed: 35720479
- Lin R et al. TRPM2 promotes pancreatic cancer by PKC/MAPK pathway. Cell Death Dis 12:585 (2021). PubMed: 34099637
- Deng D et al. Long non-coding RNA SNHG16 regulates cell behaviors through miR-542-3p/HNF4a axis via RAS/RAF/MEK/ERK signaling pathway in pediatric neuroblastoma cells. Biosci Rep 40:N/A (2020). PubMed: 32412051