Validated using a knockout cell line
Recombinant
RabMAb

Anti-B Raf antibody [EP152Y] (ab33899)

Overview

  • Product name
    Anti-B Raf antibody [EP152Y]
    See all B Raf primary antibodies
  • Description
    Rabbit monoclonal [EP152Y] to B Raf
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, IPmore details
    Unsuitable for: Flow Cyt or ICC
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. within Human B Raf aa 50-150. The exact sequence is proprietary.
    Database link: P15056

  • Positive control
    • HeLa cell lysate, human prostate cancer tissue. This antibody also reacts with rat brain tissue.
  • 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 ab33899 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
WB 1/1000 - 1/5000. Detects a band of approximately 87 kDa (predicted molecular weight: 85 kDa).
IHC-P Use at an assay dependent concentration.
IP 1/50.
  • Application notes
    Is unsuitable for Flow Cyt or ICC.
  • Target

    • Function
      Involved in the transduction of mitogenic signals from the cell membrane to the nucleus. May play a role in the postsynaptic responses of hippocampal neuron.
    • Tissue specificity
      Brain and testis.
    • Involvement in disease
      Note=Defects in BRAF are found in a wide range of cancers.
      Defects in BRAF may be a cause of colorectal cancer (CRC) [MIM:114500].
      Defects in BRAF are involved in lung cancer (LNCR) [MIM:211980].
      Defects in BRAF are involved in non-Hodgkin lymphoma (NHL) [MIM:605027]. NHL is a cancer that starts in cells of the lymph system, which is part of the body's immune system. NHLs can occur at any age and are often marked by enlarged lymph nodes, fever and weight loss.
      Defects in BRAF are a cause of cardiofaciocutaneous syndrome (CFC syndrome) [MIM:115150]; also known as cardio-facio-cutaneous syndrome. CFC syndrome is characterized by a distinctive facial appearance, heart defects and mental retardation. Heart defects include pulmonic stenosis, atrial septal defects and hypertrophic cardiomyopathy. Some affected individuals present with ectodermal abnormalities such as sparse, friable hair, hyperkeratotic skin lesions and a generalized ichthyosis-like condition. Typical facial features are similar to Noonan syndrome. They include high forehead with bitemporal constriction, hypoplastic supraorbital ridges, downslanting palpebral fissures, a depressed nasal bridge, and posteriorly angulated ears with prominent helices. The inheritance of CFC syndrome is autosomal dominant.
      Defects in BRAF are the cause of Noonan syndrome type 7 (NS7) [MIM:613706]. Noonan syndrome is a disorder characterized by facial dysmorphic features such as hypertelorism, a downward eyeslant and low-set posteriorly rotated ears. Other features can include short stature, a short neck with webbing or redundancy of skin, cardiac anomalies, deafness, motor delay and variable intellectual deficits.
      Defects in BRAF are the cause of LEOPARD syndrome type 3 (LEOPARD3) [MIM:613707]. LEOPARD3 is a disorder characterized by lentigines, electrocardiographic conduction abnormalities, ocular hypertelorism, pulmonic stenosis, abnormalities of genitalia, retardation of growth, and sensorineural deafness.
      Note=A chromosomal aberration involving BRAF is found in pilocytic astrocytomas. A tandem duplication of 2 Mb at 7q34 leads to the expression of a KIAA1549-BRAF fusion protein with a constitutive kinase activity and inducing cell transformation.
    • 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.
    • Cellular localization
      Nucleus. Cytoplasm. Cell membrane. Colocalizes with RGS14 and RAF1 in both the cytoplasm and membranes.
    • Information by UniProt
    • Database links
    • Alternative names
      • FLJ95109 antibody
      • 94 kDa B raf protein antibody
      • B raf 1 antibody
      • B Raf proto oncogene serine threonine protein kinase antibody
      • B Raf proto oncogene, serine/threonine kinase antibody
      • B RAF1 antibody
      • B-Raf proto-oncogene serine/threonine-protein kinase (p94) antibody
      • BRAF 1 antibody
      • BRAF antibody
      • BRAF_HUMAN antibody
      • BRAF1 antibody
      • cRmil antibody
      • MGC126806 antibody
      • MGC138284 antibody
      • Murine sarcoma viral (v-raf) oncogene homolog B1 antibody
      • Murine sarcoma viral v raf oncogene homolog B1 antibody
      • NS7 antibody
      • Oncogen BRAF antibody
      • oncogene BRAF1 antibody
      • p94 antibody
      • Proto-oncogene B-Raf antibody
      • Proto-oncogene c-Rmil antibody
      • RAFB 1 antibody
      • RAFB1 antibody
      • RMIL antibody
      • Serine/threonine-protein kinase B-raf antibody
      • v raf murine sarcoma viral oncogene homolog B antibody
      • v raf murine sarcoma viral oncogene homolog B1 antibody
      • v-Raf murine sarcoma viral oncogene homolog B1 antibody
      see all

    Images

    • Lane 1: Wild-type HAP1 whole cell lysate (40 µg)
      Lane 2: B Raf knockout HAP1 whole cell lysate (40 µg)
      Lane 3: HeLa whole cell lysate (40 µg)

      Lanes 1 - 3: Merged signal (red and green). Green - ab33899 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab33899 was shown to recognize B Raf in wild-type HAP1 cells as signal was lost at the expected MW in B Raf knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and B Raf knockout samples were subjected to SDS-PAGE. ab33899 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • This image shows paraffin embedded human prostate cancer tissue sample stained with ab33899 at 1/250 dilution.
    • ab33899 staining B Raf cells from human prostate tissue by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). Cells were formaldehyde fixed and permeabilized in PBS-Tween 20 prior to blocking in 70% serum for 10 minutes at 25°C. The primary antibody was diluted 1/250 and incubated with the sample for 1 hour at 25°C. A biotin conjugated goat polyclonal to mouse Ig was used as the secondary.

      See Abreview

    • Anti-B Raf antibody [EP152Y] (ab33899) at 1/5000 dilution + HeLa cell lysate

      Predicted band size: 85 kDa
      Observed band size: 87 kDa (why is the actual band size different from the predicted?)

    • All lanes : Anti-B Raf antibody [EP152Y] (ab33899) at 1/1000 dilution

      Lane 1 : Lysate prepared from mouse brain
      Lane 2 : Lysate prepared from mouse heart
      Lane 3 : Lysate prepared from mouse kidney
      Lane 4 : Lysate prepared from mouse spleen
      Lane 5 : Lysate prepared from rat brain
      Lane 6 : Lysate prepared from rat heart
      Lane 7 : Lysate prepared from rat kidney
      Lane 8 : Lysate prepared from rat spleen

      Predicted band size: 85 kDa


      Exposure time: 3 minutes

    References

    This product has been referenced in:
    • Du Z  et al. Genome-wide transcriptional analysis of BRD4-regulated genes and pathways in human glioma U251 cells. Int J Oncol N/A:N/A (2018). Read more (PubMed: 29568956) »
    • Ding Z  et al. Griffipavixanthone, a dimeric xanthone extracted from edible plants, inhibits tumor metastasis and proliferation via downregulation of the RAF pathway in esophageal cancer. Oncotarget 7:1826-37 (2016). WB . Read more (PubMed: 26646323) »

    See all 10 Publications for this product

    Customer reviews and Q&As

    Filter by Application

    Filter by Species

    Filter by Ratings

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Methanol
    Permeabilization
    No
    Username

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Sep 05 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (melanoma cell lines)
    Loading amount
    50 µg
    Specification
    melanoma cell lines
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted Jun 02 2010

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Prostate)
    Specification
    Prostate
    Fixative
    Formaldehyde
    Antigen retrieval step
    Other
    Permeabilization
    Yes - PBS-Tween20
    Blocking step
    methanol + hydrogen peroxide as blocking agent for 10 minute(s) · Concentration: 70% · Temperature: 25°C
    Username

    Dr. Serdar Balci

    Verified customer

    Submitted Nov 11 2009

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (saggital sections of cerebellum and cortex)
    Specification
    saggital sections of cerebellum and cortex
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid pH6
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
    Username

    Mr. Carl Hobbs

    Verified customer

    Submitted Feb 03 2009

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (adult brain)
    Specification
    adult brain
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid Hier, pH6
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
    Username

    Mr. Carl Hobbs

    Verified customer

    Submitted Feb 02 2009

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (brain, liver, skin from Tg animal expressing Human)
    Loading amount
    100 µg
    Specification
    brain, liver, skin from Tg animal expressing Human
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Username

    Abcam user community

    Verified customer

    Submitted Oct 02 2006

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Glioma cell line)
    Loading amount
    50 µg
    Specification
    Glioma cell line
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Username

    Abcam user community

    Verified customer

    Submitted Aug 24 2006

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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