Overview

  • Product name
    Anti-Raf1 antibody [EP4969] - N-terminal
    See all Raf1 primary antibodies
  • Description
    Rabbit monoclonal [EP4969] to Raf1 - N-terminal
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Raf1 aa 1-100. The exact sequence is proprietary.
    Database link: P04049

  • Positive control
    • HeLa and K562 cell lysates; K562 cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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.

Application Abreviews Notes
Flow Cyt 1/100.
WB 1/1000. Detects a band of approximately 73 kDa (predicted molecular weight: 73 kDa).
ICC/IF 1/100 - 1/250.

Target

  • 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
    modifications
    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.
  • Cellular localization
    Cytoplasm. Cell membrane. Colocalizes with RGS14 and BRAF in both the cytoplasm and membranes.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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
    • NS5 antibody
    • Oncogene MIL antibody
    • Oncogene RAF1 antibody
    • OTTHUMP00000160218 antibody
    • OTTHUMP00000207813 antibody
    • OTTHUMP00000209389 antibody
    • Protein kinase raf 1 antibody
    • Proto-oncogene c-RAF antibody
    • Raf 1 antibody
    • Raf 1 proto oncogene serine/threonine kinase antibody
    • RAF antibody
    • Raf proto oncogene serine/threonine protein kinase antibody
    • RAF proto-oncogene serine/threonine-protein kinase antibody
    • RAF-1 antibody
    • RAF1 antibody
    • RAF1_HUMAN antibody
    • Similar to murine leukemia viral (V-raf-1) oncogene homolog 1 antibody
    • TRANSFORMING REPLICATION-DEFECTIVE MURINE RETROVIRUS 3611-MSV antibody
    • v raf 1 murine leukemia viral oncogene homolog 1 antibody
    • v-raf murine sarcoma viral oncogene homolog 1 antibody
    • v-raf-1 murine leukemia viral oncogene-like protein 1 antibody
    • vraf1 murine leukemia viral oncogene homolog 1 antibody
    see all

Images

  • 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

    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.

  • Immunofluorescent analysis of acetone-fixed K562 cells labeling Raf1 with ab181115 at 1/250 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody at 1/200 dilution. Counter stained with Dapi.

  • Flow cytometry analysis of K-562 ( Human chronic myelogenous leukemia lymphoblast) cells labelling with 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 ab181115 and a competitor's top cited discontinued rabbit polyclonal antibody.

  • All lanes : Anti-Raf1 antibody [EP4969] - N-terminal (ab181115) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : K562 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

    Predicted band size: 73 kDa
    Observed band size: 73 kDa

References

ab181115 has not yet been referenced specifically in any publications.

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