Product nameAnti-Raf1 antibody [EP4969] - N-terminal
See all Raf1 primary antibodies
DescriptionRabbit monoclonal [EP4969] to Raf1 - N-terminal
Tested applicationsSuitable for: Flow Cyt, WB, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human Raf1 aa 1-100. The exact sequence is proprietary.
Database link: P04049
- WB: HeLa, K-562, RAW 264.7, C2C12, NIH/3T3, and PC-12 lysates. ICC/IF: HeLa cells.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Storage instructionsShipped 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 bufferPreservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab181115 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Detects a band of approximately 73 kDa (predicted molecular weight: 73 kDa).|
|ICC/IF||1/100 - 1/250.|
FunctionInvolved 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 specificityIn skeletal muscle, isoform 1 is more abundant than isoform 2.
Involvement in diseaseDefects 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 similaritiesBelongs 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.
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 localizationCytoplasm. Cell membrane. Colocalizes with RGS14 and BRAF in both the cytoplasm and membranes.
- Information by UniProt
- c Raf antibody
- C-raf antibody
- C-Raf proto-oncogene, serine/threonine kinase antibody
Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
Lanes 2, 4 and 6: Raf1 knockout HAP1 cell lysate (20 µg)
Lanes 1 and 2: Green signal from target – ab181115 observed at 74 kDa
Lanes 3 and 4: Red signal from loading control – ab8226 observed at 42 kDa
Lanes 5 and 6: Merged (red and green) signal
Unpurified ab181115 was shown to specifically react with Raf1 when Raf1 knockout samples were used. Wild-type and Raf1 knockout samples were subjected to SDS-PAGE. ab181115 and ab8226 (loading control to beta actin) were both diluted 1/1000 and incubated overnight at 4ºC. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes : Anti-Raf1 antibody [EP4969] - N-terminal (ab181115) at 1/1000 dilution ((unpurified))
Lane 1 : HeLa cell lysate
Lane 2 : K562 cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Predicted band size: 73 kDa
Observed band size: 73 kDa
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Raf1 with purified ab181115 at 1/100 dilution (10 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Flow cytometry analysis of K-562 ( Human chronic myelogenous leukemia lymphoblast) cells labelling with unpurified ab181115 at 1/100 dilution (10.79 µg/mL) (red). Cells were fixed with 4% paraformaldehyde . Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/2000 dilution. Isotype control - 90% methanol . Unlabeled control - Rabbit monoclonal IgG (ab172730) / Black.
Lanes 1: Wild-type HAP1 cell lysate (20 µg)
Lanes 2: Raf1 knockout HAP1 cell lysate (20 µg)
Lanes 1 and 2: Merged signal (red and green).
Green - Target observed at 74 kDa. Red - loading control, ab8226, observed at 42 kDa
This western blot image is a comparison between unpurified ab181115 and a competitor's top cited discontinued rabbit polyclonal antibody.
Immunofluorescent analysis of acetone-fixed K562 cells labeling Raf1 with unpurified ab181115 at 1/250 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody at 1/200 dilution. Counter stained with Dapi.
ab181115 has not yet been referenced specifically in any publications.