Anti-IDH2 antibody [5F11]

7 product images

• 

  Western blot - Anti-IDH2 antibody [5F11] (ab55271)

  Western blot against tagged recombinant protein immunogen using ab55271 IDH2 antibody at 1ug/ml. Predicted band size of immunogen is 37 kDa
  

  This image was generated using the ascites version of the product.

  All lanes: Western blot - Anti-IDH2 antibody [5F11] (ab55271)

  Predicted band size: 50 kDa

• 

  Flow Cytometry - Anti-IDH2 antibody [5F11] (ab55271)

  Overlay histogram showing MCF7 cells stained with ab55271 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab55271, 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 antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  
  

  This image was generated using the ascites version of the product.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH2 antibody [5F11] (ab55271)

  IDH2 antibody (ab55271) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human colon.
  

  This image was generated using the ascites version of the product.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-IDH2 antibody [5F11] (ab55271)

  ICC/IF image of ab55271 stained Mcf7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab55271, 10μg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.
  
  

  This image was generated using the ascites version of the product.

• 

  Western blot - Anti-IDH2 antibody [5F11] (ab55271)

  The blocking agent used is 5% milk.
  

  This image was generated using the ascites version of the product.

  All lanes: Western blot - Anti-IDH2 antibody [5F11] (ab55271)

  Lane 1: IDH2 in transfected 293T cell line

  Lane 2: Non-transfected lysate

  Predicted band size: 50 kDa

• 

  Western blot - Anti-IDH2 antibody [5F11] (ab55271)

  False colour image of Western blot: Anti-IDH2 antibody [5F11] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab55271 was shown to bind specifically to IDH2. A band was observed at 48 kDa in wild-type Jurkat cell lysates with no signal observed at this size in IDH2 knockout cell line Human IDH2 knockout Jurkat cell line ab282331 (knockout cell lysate ab283148). To generate this image, wild-type and IDH2 knockout Jurkat 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-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.

  All lanes: Western blot - Anti-IDH2 antibody [5F11] (ab55271) at 1/1000 dilution

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

  Lane 2: IDH2 knockout Jurkat cell lysate at 20 µg

  Lane 2: Western blot - Human IDH2 knockout Jurkat cell line (Human IDH2 knockout Jurkat cell line ab282331)

  Lane 3: MOLT-4 cell lysate at 20 µg

  Lane 4: HEK-293 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 50 kDa

  Observed band size: 48 kDa

• 

  Western blot - Anti-IDH2 antibody [5F11] (ab55271)

  Western blot: Anti-IDH2 antibody [5F11] (ab55271) staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab55271 was shown to bind specifically to IDH2. A band was observed at 51 kDa in wild-type A549 cell lysates with no signal observed at this size in IDH2 knockout cell line. To generate this image, wild-type and IDH2 knockout A549 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-IDH2 antibody [5F11] (ab55271) at 1/1000 dilution

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: IDH2 knockout A549 cell lysate at 20 µg

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

  Lane 4: IDH2 knockout Jurkat Human IDH2 knockout Jurkat cell line ab282331 cell lysate at 20 µg

  Secondary

  All lanes: Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 51 kDa