Anti-MiTF antibody [D5] - BSA and Azide free

3 product images

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  Immunocytochemistry - Anti-MiTF antibody [D5] - BSA and Azide free (ab182842)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-MiTF antibody [D5] ab3201).

  Anti-MiTF antibody [D5] ab3201 staining MiTF in Malme-3M cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-MiTF antibody [D5] ab3201 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

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  Flow Cytometry (Intracellular) - Anti-MiTF antibody [D5] - BSA and Azide free (ab182842)

  Overlay histogram showing Malme-3 cells stained with Anti-MiTF antibody [D5] ab3201 (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. The cells were then incubated with the antibody (Anti-MiTF antibody [D5] ab3201, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2μg/1x10⁶ cells ) used under the same conditions. Acquisition of >5,000 events was performed.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-Arginine and sodium azide (Anti-MiTF antibody [D5] ab3201).

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  Western blot - Anti-MiTF antibody [D5] - BSA and Azide free (ab182842)

  This data was developed using ab127540, the same antibody clone in a different buffer formulation.
  

  7.5% SDS-PAGE

  All lanes: Anti-ZNF282 antibody (ab127540) at 1/1000 dilution

  All lanes: Raji whole cell lysate at 30 µg

  Predicted band size: 58 kDa