Anti-Dnmt1 antibody [EPR3522] - BSA and Azide free

6 product images

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  Western blot - Anti-Dnmt1 antibody [EPR3522] - BSA and Azide free (ab207601)

  This WB data was generated using the same anti-Dnmt1 antibody clone, EPR3522, in a different buffer formulation (cat# Anti-Dnmt1 antibody [EPR3522] ab92314).
  

  Lane 1: Wild type HAP1 whole cell lysate (20 μg)
  Lane 2: DNMT1 knockout HAP1 whole cell lysate (20 μg)
  Lane 3: HEK293 whole cell lysate (20 μg)
  Lane 4: HeLa whole cell lysate (20 μg)
  Lanes 1 - 4: Merged signal (red and green). Green - Anti-Dnmt1 antibody [EPR3522] ab92314 observed at 170 kDa. Red - loading control, ab18058, observed at 130 kDa.
  
  

  Anti-Dnmt1 antibody [EPR3522] ab92314 was shown to specifically react with DNMT1 when DNMT1 knockout samples were used. Wild-type and DNMT1 knockout samples were subjected to SDS-PAGE. Anti-Dnmt1 antibody [EPR3522] ab92314 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-Dnmt1 antibody [EPR3522] (Anti-Dnmt1 antibody [EPR3522] ab92314)

  Predicted band size: 183 kDa

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  Flow Cytometry (Intracellular) - Anti-Dnmt1 antibody [EPR3522] - BSA and Azide free (ab207601)

  Overlay histogram showing HeLa cells stained with Anti-Dnmt1 antibody [EPR3522] ab92314 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Anti-Dnmt1 antibody [EPR3522] ab92314, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Dnmt1 antibody [EPR3522] ab92314).
  

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  Immunoprecipitation - Anti-Dnmt1 antibody [EPR3522] - BSA and Azide free (ab207601)

  This data was developed using Anti-Dnmt1 antibody [EPR3522] ab92314, the same antibody clone in a different buffer formulation.Dnmt1 was immunoprecipitated from 0.35 mg Hut-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate 10 µg with Anti-Dnmt1 antibody [EPR3522] ab92314 at 1/50 dilution (2µg). VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

  Lane 1: Hut-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate 10 µg

  Lane 2: abab92314 IP in Hut-78 whole cell lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Dnmt1 antibody [EPR3522] ab92314 in Hut-78 whole cell lysate

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

  All lanes: Immunoprecipitation - Anti-Dnmt1 antibody [EPR3522] (Anti-Dnmt1 antibody [EPR3522] ab92314)

  Predicted band size: 183 kDa

  Observed band size: 183 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-Dnmt1 antibody [EPR3522] - BSA and Azide free (ab207601)

  This ICC/IF data was generated using the same anti-Dnmt1 antibody clone, EPR3522, in a different buffer formulation (cat# Anti-Dnmt1 antibody [EPR3522] ab92314).
  

  Immunofluorescence staining of Jurkat cells with purified Anti-Dnmt1 antibody [EPR3522] ab92314 at a working dilution of 1/2000, counter-stained with DAPI. The secondary antibody was an Alexa Fluor^® 488 conjugated goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt1 antibody [EPR3522] - BSA and Azide free (ab207601)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Dnmt1 antibody [EPR3522] ab92314).

  Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Dnmt1 with Anti-Dnmt1 antibody [EPR3522] ab92314 at 1/200 (3.55 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on human tonsil. The section was incubated with Anti-Dnmt1 antibody [EPR3522] ab92314 for 30 mins at room temperature.
  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

  Counterstained with Hematoxylin.

  Secondary antibody only control: Ready to use LeicaDS9800 (Bond™ Polymer Refine Detection)

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt1 antibody [EPR3522] - BSA and Azide free (ab207601)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Dnmt1 antibody [EPR3522] ab92314).

  Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling Dnmt1 with Anti-Dnmt1 antibody [EPR3522] ab92314 at 1/200 (3.55 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on human placenta. The section was incubated with Anti-Dnmt1 antibody [EPR3522] ab92314 for 30 mins at room temperature.
  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

  Counterstained with Hematoxylin.

  Secondary antibody only control: Ready to use LeicaDS9800 (Bond™ Polymer Refine Detection)

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.