Phospho-Raf1 (S621) Recombinant Rabbit Monoclonal Antibody [JJ085-05]
Catalog# ET1701-3
Phospho-Raf1 (S621) Recombinant Rabbit Monoclonal Antibody [JJ085-05]
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WB
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IHC-P
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Human
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Rat
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Mouse
Predicted species support after-sales service
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unconjugated
Overview
Product Name
Phospho-Raf1 (S621) Recombinant Rabbit Monoclonal Antibody [JJ085-05]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding Ser621 of Human Raf1 aa 591-640 / 648.
Species Reactivity
Human, Rat (Predicted: Mouse)
Predicted species support after-sales service
Validated Applications
WB, IHC-P
Molecular Weight
Predicted band size: 73 kDa
Positive Control
Human skeletal muscle tissue lysates, rat trachea tissue, human muscle tissue, human kidney tissue.
Conjugation
unconjugated
Clone Number
JJ085-05
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:2,000
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IHC-P
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1:100-1:500
Target
Function
Raf-1 is a ubiquitously expressed cytoplasmic protein with intrinsic serine/threonine kinase activity. Raf-1, or c-Raf, is the cellular homolog of v-Raf, the product of the transforming gene of the 3611 strain of murine sarcoma virus. The unregulated kinase activity of the v-Raf protein is associated with cellular transformation and mitogenesis. Raf-1 is normally suppressed by its regulatory N-terminal domain. Raf-1 is activated in response to a variety of tyrosine kinase receptors as well as in response to pp60v-Src expression. Specifically, Raf-1 is phosphorylated in the catalytic domain at Ser 338 and, to a lesser extent, Ser 339. This phosphorylation requires the co-activation of PI 3-kinase and the Ras signaling pathway. Raf-1 is also phosphorylated on Tyr 340 and 341, which induces the phosphorylation of MEK. Phosphorylation of Ser 621 is essential for the catalytic activity of Raf-1 and downregulation by c-AMP-dependent protein kinase A (PKA). PKA also phosphorylates Raf-1 on Ser 43 and Ser 259. PKA phosphorylation of Ser 259 inhibits Raf-1 and decreases the phosphorylation necessary for Raf-1 activation at Ser 338.
Background References
1. K hler, M. et al. 2016. Activation loop phosphorylation regulates B-Raf in vivo and transformation by B-Raf mutants. The EMBO journal. 35: 143-61.
2. El-Chaar, NN. et al. 2014. Genomic classification of the RAS network identifies a personalized treatment strategy for lung cancer. Molecular oncology. 8: 1339-54.
Sequence Similarity
Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. RAF subfamily.
Tissue Specificity
In skeletal muscle, isoform 1 is more abundant than isoform 2.
Post-translational Modification
Phosphorylation at Thr-269, Ser-338, Tyr-341, Thr-491 and Ser-494 results in its activation. Phosphorylation at Ser-29, Ser-43, Ser-289, Ser-296, Ser-301 and Ser-642 by MAPK1/ERK2 results in its inactivation. 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. Phosphorylation at Ser-338 by PAK1 and PAK5 and Ser-339 by PAK1 is required for its mitochondrial localization. Phosphorylation at Ser-621 in response to growth factor treatment stabilizes the protein, possibly by preventing proteasomal degradation. Phosphorylation at Ser-289, Ser-296, Ser-301, Ser-338 and Ser-621 are somehow linked to the methylation potential of cells. Treatment of cells with HGF in the presence of the methylation inhibitor 5'-methylthioadenosine (MTA) results in increased phosphorylation at Ser-338 and Ser-621 and decreased phosphorylation at Ser-296, Ser-301 and Ser-338. Dephosphorylation at Ser-338 by PPP5C results in an activity decrease.; Methylated at Arg-563 in response to EGF treatment. This modification leads to destabilization of the protein, possibly through proteasomal degradation.
Subcellular Location
Cytoplasm, Cell membrane, Mitochondrion, Nucleus.
Synonyms
c Raf antibody
C-raf antibody
C-Raf proto-oncogene, serine/threonine kinase antibody
CMD1NN antibody
Craf 1 transforming gene antibody
cRaf antibody
Craf1 transforming gene antibody
EC 2.7.11.1 antibody
kinase Raf1 antibody
Murine sarcoma 3611 oncogene 1 antibody
ExpandImages
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Western blot analysis of Phospho-Raf1 (S621) on human skeletal muscle tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-3, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
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Immunohistochemical analysis of paraffin-embedded rat trachea tissue using anti-Phospho-Raf1 (S621) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-3, 1/400) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human muscle tissue using anti-Phospho-Raf1 (S621) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-3, 1/400) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Phospho-Raf1 (S621) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-3, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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