Overview

  • Product nameAnti-B Raf antibody [EP152Y]See all B Raf primary antibodies ...
  • Description
    Rabbit monoclonal [EP152Y] to B Raf
  • Tested applicationsWB, IHC-P, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide:

    CGEHNPPSIYLEAYEEYT

    , corresponding to N terminal amino acids 70-86 of Human B Raf

  • Positive control
    • HeLa cell lysate, human prostate cancer tissue. This antibody also reacts with rat brain tissue.
  • General notesProduced under U.S. Patent No. 5,675,063.

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.
Flow Cyt 1/1000.
IP 1/50.
  • Application notesIs unsuitable for ICC.
  • Target

    • FunctionInvolved 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 specificityBrain and testis.
    • Involvement in diseaseNote=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 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.
    • Cellular localizationNucleus. 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 1 antibody
      • B Raf proto oncogene serine threonine protein kinase antibody
      • B Raf proto oncogene serine threonine protein kinase antibody
      • B-Raf proto-oncogene serine/threonine-protein kinase (p94) antibody
      • BRAF 1 antibody
      • BRaf antibody
      • BRaf antibody
      • BRAF_HUMAN antibody
      • BRAF1 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
      • Murine sarcoma viral v raf oncogene homolog B1 antibody
      • Murine sarcoma viral v raf oncogene homolog B1 antibody
      • NS7 antibody
      • oncogene BRAF1 antibody
      • p94 antibody
      • p94 antibody
      • Proto-oncogene B-Raf antibody
      • Proto-oncogene c-Rmil antibody
      • RAFB 1 antibody
      • RAFB1 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
      • v-Raf murine sarcoma viral oncogene homolog B1 antibody
      see all

    Anti-B Raf antibody [EP152Y] images

    • Overlay histogram showing SH-SY5Y cells stained with ab33899 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab33899, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • 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?)
    • This image shows paraffin embedded human prostate cancer tissue sample stained with ab33899 at 1/250 dilution.This image shows paraffin embedded human prostate cancer tissue sample stained with ab33899 at 1/250 dilution.
    • 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
    • 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

    References for Anti-B Raf antibody [EP152Y] (ab33899)

    This product has been referenced in:
    • Hirschi B & Kolligs FT Alternative splicing of BRAF transcripts and characterization of C-terminally truncated B-Raf isoforms in colorectal cancer. Int J Cancer N/A:N/A (2013). WB ; Human . Read more (PubMed: 23354951) »
    • Khalili JS  et al. Oncogenic BRAF(V600E) promotes stromal cell-mediated immunosuppression via induction of interleukin-1 in melanoma. Clin Cancer Res 18:5329-40 (2012). Flow Cyt ; Human . Read more (PubMed: 22850568) »

    See all 5 Publications for this product

    Product Wall

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has cau...

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

    Dr. Kirk McManus

    Verified customer

    Submitted Sep 05 2012

    Unfortunately shipment of this order is being held up by our accounts team.

    Our accounting department can be contacted by email at mailto:us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corne...

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    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

    Thank you for your enquiry. I have been in touch with the lab and I can tell you that this antibody has NOT been tested for its capacity to differentiate between wild type and variant (V600E) BRAF. I could not find literature relating to the varian...

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    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"