Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-KDM6A / UTX antibody [EPR23203-211] - BSA and Azide free (ab275402)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23203-211] to KDM6A / UTX - BSA and Azide free
  • Suitable for: WB, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-KDM6A / UTX antibody [EPR23203-211] - BSA and Azide free
    See all KDM6A / UTX primary antibodies

  • Description

    Rabbit monoclonal [EPR23203-211] to KDM6A / UTX - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt (Intra)more details
    Unsuitable for: ChIP,ICC/IF,IHC-P or IP

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

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

  • Positive control

    • WB: HAP1, HEK-293T, NIH/3T3 and F9 whole cell lysates. Flow Cyt (intra): HAP1 and F9 cells.

  • General notes

    ab275402 is the carrier-free version of ab253183.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

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

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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.

Properties

Images

  • Western blot - Anti-KDM6A / UTX antibody [EPR23203-211] - BSA and Azide free (ab275402)
    Western blot - Anti-KDM6A / UTX antibody [EPR23203-211] - BSA and Azide free (ab275402)
    All lanes : Anti-KDM6A / UTX antibody [EPR23203-211] (ab253183) at 1/1000 dilution ((0.502 µg/ml))

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : KDM6A knockout HAP1 whole cell lysate
    Lane 3 : F9 (mouse embryonal carcinoma epithelial cell), whole cell lysate
    Lane 4 : THP-1 (human monocytic leukemia monocyte), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/20000 dilution

    Predicted band size: 154 kDa
    Observed band size: 160 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab253183, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

    Lanes 1-4: Merged signal (red and green). Green - ab253183 observed at 160 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.

    ab253183 Anti-KDM6A  antibody [EPR23203-211] was shown to react with KDM6A in  HAP1 wild-type cells in Western blot. Loss of signal was observed when KDM6A knockout sample was used. Wild-type and KDM6A knockout samples were subjected to SDS-PAGE.

    ab253183 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1/1000 dilution and 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.

    Negative control: THP-1 (PMID: 19330029).

    The expression profile and molecular weight observed is consistent with what has been described in the literature (PMID: 19330029).

    The the bands between 50kDa and 150kDa are caused by degradation based on the result of fresh lysates.

  • Flow Cytometry - Anti-KDM6A / UTX antibody [EPR23203-211] - BSA and Azide free (ab275402)
    Flow Cytometry - Anti-KDM6A / UTX antibody [EPR23203-211] - BSA and Azide free (ab275402)

    This data was developed using ab253183, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized THP-1 (Human monocytic leukemia monocyte), (Left panel) / HAP1 (Human chronic myelogenous leukemia near-haploid cell line), (Right panel) cells labelling KDM6A / UTX with ab253183 at 1/50 dilution (1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730)  isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    Negative control: THP-1.

  • Western blot - Anti-KDM6A / UTX antibody [EPR23203-211] - BSA and Azide free (ab275402)
    Western blot - Anti-KDM6A / UTX antibody [EPR23203-211] - BSA and Azide free (ab275402)
    All lanes : Anti-KDM6A / UTX antibody [EPR23203-211] (ab253183) at 1/1000 dilution ((0.502 µg/ml))

    Lane 1 : HEK-293T (human embryonic kidney epithelial cell), whole cell lysate
    Lane 2 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 154 kDa
    Observed band size: 160 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Fresh lysates were used in this WB.

    Exposure time: 3 minutes.

     

  • Flow Cytometry - Anti-KDM6A / UTX antibody [EPR23203-211] - BSA and Azide free (ab275402)
    Flow Cytometry - Anti-KDM6A / UTX antibody [EPR23203-211] - BSA and Azide free (ab275402)

    This data was developed using ab253183, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized F9 (Mouse embryonal carcinoma epithelial cell) cells labelling KDM6A / UTX with ab253183 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730)  isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Anti-KDM6A / UTX antibody [EPR23203-211] - BSA and Azide free (ab275402)
    Anti-KDM6A / UTX antibody [EPR23203-211] - BSA and Azide free (ab275402)

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