Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free

5 product images

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  Western blot - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)

  This WB data was generated using the same anti-KMT6/EZH2 antibody clone, EPR9307(2), in a different buffer formulation (cat# Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080).
  

  Lane 1: Wild-type HAP1 cell lysate (20 μg)
  Lane 2: KMT6 / EZH2 knockout HAP1 cell lysate (20 μg)
  Lane 3: HEK293 cell lysate (20 μg)
  Lane 4: Ms testis tissue lysate (20 μg)
  Lanes 1 - 4: Merged signal (red and green). Green - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080 observed at 93 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

  Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080 was shown to recognize KMT6 / EZH2 when KMT6 / EZH2 knockout samples were used, along with additional cross-reactive bands. Wild-type and KMT6 / EZH2 knockout samples were subjected to SDS-PAGE. Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted at 1/500 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  All lanes: Western blot - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal (Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080)

  Predicted band size: 85 kDa, 92 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)

  Immunofluorescent analysis of HeLa cells (4% Paraformaldehyde-fixed; 0.1% tritonX-100-permeabilized) labeling KMT6 / EZH2 with Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080 at 1/250 dilution followed by Goat anti rabbit IgG (AlexaFluor® 488) secondary at 1/200 dilution and counter-stained with DAPI (blue). 

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)

  This IHC data was generated using the same anti-KMT6/EZH2 antibody clone, EPR9307(2), in a different buffer formulation (cat# Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080).
  

  Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling KMT6 / EZH2 with Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080 at 1/250 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin (inset: negative control).

  Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Flow Cytometry (Intracellular) - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)

  Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080 staining KMT6 / EZH2 in the human cell line Jurkat (human acute T cell leukemia) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/90. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.
  

  Isoytype control: Rabbit monoclonal IgG (Black)

  Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080).

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  Western blot - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - BSA and Azide free (ab231165)

  False colour image of Western blot: Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080 was shown to bind specifically to KMT6 / EZH2. A band was observed at 90 kDa (green arrow) in wild-type MCF7 cell lysates with no signal observed at this size in ezh2 CRISPR-Cas9 edited cell line ab281611 (CRISPR-Cas9 edited cell lysate ab282963). The band observed in the CRISPR-Cas9 edited lysate lane below 90 kDa (yellow arrow) is likely to represent a truncated form of KMT6 / EZH2. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and ezh2 CRISPR-Cas9 edited MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal (Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal ab191080) at 1/1000 dilution

  Lane 1: Wild-type MCF7 cell lysate at 20 µg

  Lane 2: ezh2 CRISPR-Cas9 edited MCF7 cell lysate at 20 µg

  Lane 3: HEK-293 cell lysate at 20 µg

  Lane 4: Mouse Testis cell lysate at 20 µg

  Performed under reducing conditions.

  Observed band size: 90 kDa