Rabbit monoclonal [EPR3433(2)] to Raf1 (phospho S259)
This antibody only detects Raf1 when phosphorylated at Serine 259.
WB, IHC-P, Dot blotmore details Unsuitable for:
Flow Cyt,ICC/IF or IP
Mouse, Rat, Human
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Raf1 aa 250-350 (phospho S259) (Cysteine residue). The exact sequence is proprietary. Database link: P04049
WB: HEK293 whole cell lysate.
IHC-P: Human endometrial carcinoma and human skeletal muscle tissues.
This product is a recombinant rabbit monoclonal antibody.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
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.
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use at an assay dependent concentration.
Is unsuitable for Flow Cyt,ICC/IF or IP.
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.
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.
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.
Phosphorylated 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.
Cytoplasm. Cell membrane. Colocalizes with RGS14 and BRAF in both the cytoplasm and membranes.
Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539)
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 Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size : 73 kDa Observed band size : 73 kDa
Immunohistochemical analysis of paraffin-embedded Human endometrial carcinoma tissue labeling Raf1 with ab173539 at a 1/50 dilution.
Dot Blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539)
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.