Anti-Vimentin antibody [SP20], prediluted

4 product images

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  Western blot - Anti-Vimentin antibody [SP20], prediluted (ab27608)

  This image was generated using a previous batch manufactured using the hybridoma production method.

  
  This data was developed using the same antibody clone in a different buffer formulation (Anti-Vimentin antibody [SP20] ab16700).
  

  Lanes 1 - 4: Merged signal (red and green). Green - Anti-Vimentin antibody [SP20] ab16700 observed at 53 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.

  Anti-Vimentin antibody [SP20] ab16700 was shown to react with Vimentin in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human VIM (Vimentin) knockout HeLa cell line ab255446 (knockout cell lysate Human VIM (Vimentin) knockout HeLa cell lysate ab263775) was used. Wild-type and Vimentin knockout samples were subjected to SDS-PAGE. Anti-Vimentin antibody [SP20] ab16700 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 100 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  Lanes 1 - 4: Western blot - Anti-Vimentin antibody [SP20], prediluted (ab27608) at 1/100 dilution

  Lanes 1 - 4: Western blot - Anti-Vimentin antibody [SP20] (Anti-Vimentin antibody [SP20] ab16700) at 1/100 dilution

  Lane 1: U20S cell lysate at 20 µg

  Lane 2: Human tonsil cell lysate at 20 µg

  Lane 2: Western blot - Human VIM (Vimentin) knockout HeLa cell line (Human VIM (Vimentin) knockout HeLa cell line ab255446)

  Lane 3: Wild-type HeLa cell lysate at 20 µg

  Lane 4: VIM knockout HeLa cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution

  Performed under reducing conditions.

  Predicted band size: 53 kDa

  Observed band size: 37 kDa, 53 kDa

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  Western blot - Anti-Vimentin antibody [SP20], prediluted (ab27608)

  This image refers to the non-prediluted version of this product (ab16900)
  

  This image was generated using a previous batch manufactured using the hybridoma production method.

  Lane 1: Anti-MDM2 antibody [4B11] (ab16900) at 1/100 dilution

  Lane 1: Western blot - Anti-Vimentin antibody [SP20] (Anti-Vimentin antibody [SP20] ab16700) at 1/100 dilution

  All lanes: HeLa cell lysate

  Predicted band size: 53 kDa

  Observed band size: 53 kDa

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  Flow Cytometry - Anti-Vimentin antibody [SP20], prediluted (ab27608)

  Overlay histogram showing HeLa cells stained with ab27608 (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 (ab27608, neat) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  
  

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [SP20], prediluted (ab27608)

  Human melanoma stained with prediluted ab27608.