Overview

  • Product nameAnti-B Raf antibodySee all B Raf primary antibodies ...
  • Description
    Rabbit polyclonal to B Raf
  • Tested applicationsICC/IF, WB, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Cow, Dog, Chimpanzee, Gorilla
  • Immunogen

    Synthetic peptide, corresponding to a region between residue 25 and 75 of human B Raf (NP_004324.2)

  • Positive control
    • HeLa and 293T whole cell lysate IF/ICC: MCF7 cell line.

Properties

Applications

Our Abpromise guarantee covers the use of ab85972 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
ICC/IF Use a concentration of 5 µg/ml.
WB 1/2000 - 1/10000. Predicted molecular weight: 85 kDa.
IP Use at 2-5 µg/mg of lysate.
IHC-P 1/200 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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 proto oncogene serine threonine protein kinase 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
    • p94 antibody
    • Proto-oncogene B-Raf antibody
    • Proto-oncogene c-Rmil antibody
    • RAFB 1 antibody
    • RAFB1 antibody
    • Serine/threonine-protein kinase B-raf 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 images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling B Raf with ab85972 at 1/200 (1µg/ml). Detection: DAB.
  • ICC/IF image of ab85972 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab85972, 1/200 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM

  • All lanes : Anti-B Raf antibody (ab85972) at 0.04 µg/ml

    Lane 1 : HeLa whole cell lysate at 50 µg
    Lane 2 : HeLa whole cell lysate at 15 µg
    Lane 3 : HeLa whole cell lysate at 5 µg
    Lane 4 : 293T whole cell lysate at 50 µg


    Predicted band size : 85 kDa
    Observed band size : 95 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 238 kDa,60 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes
  • Detection of B Raf by Western Blot of Immunprecipitate.
    ab85972 at 1µg/ml staining B Raf in HeLa whole cell lysate immunoprecipitated using ab85972 at 3µg/mg lysate (1 mg/IP; 20% of IP loaded/lane). Detection: Chemiluminescence with exposure time of 10 seconds.

References for Anti-B Raf antibody (ab85972)

ab85972 has not yet been referenced specifically in any publications.

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