Anti-Cyclin D2 antibody [EPR27468-62]

4 product images

• 

  Western blot - Anti-Cyclin D2 antibody [EPR27468-62] (ab308258)

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

  Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as a GAPDH loading control.

  We recommend using a higher sensitive ECL substrate to increase the band intensity.

  All lanes: Western blot - Anti-Cyclin D2 antibody [EPR27468-62] (ab308258) at 1/1000 dilution

  Lane 1: 293T (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg

  Lane 2: U-2 OS (Human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: Human heart tissue lysate at 20 µg

  Lane 4: Human thyroid tissue lysate at 20 µg

  Lane 5: Mouse heart tissue lysate at 20 µg

  Lane 6: Mouse thyroid tissue lysate at 20 µg

  Lane 7: Rat heart tissue lysate at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Developed using the ECL technique.

  Predicted band size: 33 kDa

  Observed band size: 31 kDa

  Exposure time: 40s

• 

  Western blot - Anti-Cyclin D2 antibody [EPR27468-62] (ab308258)

  Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBS. Lysates at 20 µg per lane.
  
  The samples were run on a Bis-Tris gel under reducing conditions.
  
  Western blot: Anti-Cyclin D2 antibody (ab308258) staining at 1/1000 dilution, shown in green; Mouse anti-Tubulin antibody DM1A (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red.
  
  In Western blot, ab308258 was shown to bind specifically to  Cyclin D2. Target of interest was observed at 31 kDa in wild-type 293T cell lysates (lane 1) with no signal observed at this size in Cyclin D2 knockout cell line Human CCND2 (Cyclin D2) knockout HEK-293T cell line ab267318(Cyclin D2 knockout cell lysate Human CCND2 (Cyclin D2) knockout HEK-293T cell lysate ab257875) (lane 2).  To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane.  Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C.  Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

  All lanes: Western blot - Anti-Cyclin D2 antibody [EPR27468-62] (ab308258) at 1/1000 dilution

  Lane 1: Wild-type 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

  Lane 2: CCND2 knockout 293T whole cell lysate at 20 µg

  Lane 3: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

  Lane 4: C2C12 (mouse myoblast) whole cell lysate at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG H&L (800CW) and Goat Anti-Mouse IgG H&L (680RD) at 1/10000 dilution

  Predicted band size: 32 kDa

  Observed band size: 31 kDa

• 

  Western blot - Anti-Cyclin D2 antibody [EPR27468-62] (ab308258)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  This antibody does not cross-react with mouse CCND3 and CCND1.
  
  In Western blot, anti-His antibody (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.#8 seconds
  
  Exposure time: 8 seconds

  All lanes: Western blot - Anti-Cyclin D2 antibody [EPR27468-62] (ab308258) at 1/1000 dilution

  Lane 1: His-tagged mouse CCND2 recombinant protein fragment at 10 ng

  Lane 2: His-tagged mouse CCND3 recombinant protein at 10 ng

  Lane 3: MYC/DDK -tagged mouse CCND1 recombinant protein at 10 ng

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 32 kDa

  Observed band size: 31 kDa

  Exposure time: 8s

• 

  Western blot - Anti-Cyclin D2 antibody [EPR27468-62] (ab308258)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] ab129002) loading control staining at 1/10000 dilution.
  
  Exposure time: 180 seconds

  All lanes: Western blot - Anti-Cyclin D2 antibody [EPR27468-62] (ab308258) at 1/1000 dilution

  Lane 1: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: RAW 264.7 (mouse elson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

  Lane 3: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

  Lane 4: Mouse spleen tissue lysate at 20 µg

  Lane 5: Rat spleen tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 32 kDa

  Observed band size: 31 kDa

  Exposure time: 180s