Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP152Y] to BRAF - BSA and Azide free
  • Suitable for: WB, IHC-P, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-BRAF antibody [EP152Y] - BSA and Azide free
    See all BRAF primary antibodies

  • Description

    Rabbit monoclonal [EP152Y] to BRAF - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

     

  • Tested applications

    Suitable for: WB, IHC-P, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa (ab150035) and HAP1 whole cell lysate. IHC-P: human prostate cancer tissue. This antibody also reacts with rat brain tissue. IP: HeLa cells.

  • General notes

    Ab189351 is the carrier-free version of ab33899. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab189351 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)
    Western blot - Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)
    All lanes : Anti-BRAF antibody [EP152Y] (ab33899) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : BRAF knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 85 kDa
    Observed band size: 90 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab33899).

      Lanes 1- 2: Merged signal (red and green). Green - ab33899 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab33899 was shown to react with BRAF in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265373 (knockout cell lysate ab257078) was used. Wild-type HeLa and BRAF knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab33899 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling BRAF with purified ab33899 at 1/100 dilution (4.77 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33899).

  • Western blot - Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)
    Western blot - Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)
    All lanes : Anti-BRAF antibody [EP152Y] (ab33899) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : BRAF knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate

    Lysates/proteins at 40 µg per lane.

    Predicted band size: 85 kDa



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

    ab33899 was shown to recognize BRAF in wild-type HAP1 cells as signal was lost at the expected MW in BRAF knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and BRAF 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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33899). 

  • Immunoprecipitation - Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)
    Immunoprecipitation - Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)

    ab33899 (Purified) at 1:500 dilution (0.954 µg/ml) immunoprecipitating BRAF in HeLa whole cell lysate .
    Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
    Lane 2 (+): ab33899 & HeLa whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab33899 in HeLa whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM /TBST .This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189351)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)

    This IHC data was generated using the same anti-B Raf antibody clone, EP152Y, in a different buffer formulation (cat# ab33899).

    ab33899 (unpurified) 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)

    This IHC data was generated using the same anti-B Raf antibody clone, EP152Y, in a different buffer formulation (cat# ab33899).

    This image shows paraffin embedded human prostate cancer tissue sample stained with ab33899 (unpurified) at 1/250 dilution.

  • Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)
    Anti-BRAF antibody [EP152Y] - BSA and Azide free (ab189351)

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