Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP592Y] to TRAF6 - BSA and Azide free
  • Suitable for: WB, ICC/IF, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-TRAF6 antibody [EP592Y] - BSA and Azide free
    See all TRAF6 primary antibodies

  • Description

    Rabbit monoclonal [EP592Y] to TRAF6 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HAP1, Daudi, Jurkat, HEK293 and HeLa cell lysates. IHC-P: Human cerebral cortex and mouse kidney tissues. ICC/IF: HeLa cells.

  • General notes

    Ab227560 is the carrier-free version of ab40675. 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

    ab227560 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)
    Western blot - Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)
    All lanes : Anti-TRAF6 antibody [EP592Y] (ab40675) at 1/500 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : TRAF6 knockout HeLa cell lysate
    Lane 3 : HAP1 cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 63 kDa
    Observed band size: 65 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 (ab40675).

    Lanes 1- 4: Merged signal (red and green). Green - ab40675 observed at 65 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab40675 was shown to react with TRAF6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab266009 (knockout cell lysate ab257760) was used. Wild-type HeLa and TRAF6 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. ab40675 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 500 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-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labelling TRAF6 with purified ab40675 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40675).

  • Immunocytochemistry/ Immunofluorescence - Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)
    Immunocytochemistry/ Immunofluorescence - Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)

    Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling TRAF6 with purified ab40675 at 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/50) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40675).

  • Western blot - Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)
    Western blot - Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)

    This WB data was generated using the same anti-TRAF6 antibody clone, EP592Y, in a different buffer formulation (cat# ab40675).

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: TRAF6 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: HEK293 whole cell lysate (20 µg)

     

    Lanes 1 - 4: Merged signal (red and green). Green - ab40675 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.

     

    Ab40675 was shown to specifically react with TRAF6 in wild-type cells, along with additional cross-reactive bands as signal was lost in TRAF6 knockout HAP1 cells. Wild-type and TRAF6 knockout samples were subjected to SDS-PAGE. Ab40675 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)
    Immunocytochemistry/ Immunofluorescence - Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)

    Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling TRAF6 with unpurified ab40675 at 1/250.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40675).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)

    This IHC data was generated using the same anti-TRAF6 antibody clone, EP592Y, in a different buffer formulation (cat# ab40675).

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebral cortex tissue labelling TRAF6 with purified ab40675 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)
    Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)

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