Anti-p27 KIP 1 antibody [SX53G8]

4 product images

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  Immunocytochemistry - Anti-p27 KIP 1 antibody [SX53G8] (ab193379)

  Immunocytochemical analysis of PFA-fixed MCF-7 cells labeling p27 KIP 1 with ab193379 at 1 μg/mL. A Goat anti-mouse IgG CF488 antibody (Green) was used as a secondary antibody. The membrane is stained with Phalloidin-CF640

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  Western blot - Anti-p27 KIP 1 antibody [SX53G8] (ab193379)

  Lane 1: Wild-type HAP1 whole cell lysate (20 μg)
  Lane 2: CDKN1B knockout HAP1 whole cell lysate (20 μg)
  Lane 3: HeLa whole cell lysate (20 μg)
  Lane 4: MCF7 whole cell lysate (20 μg)
  

  Lanes 1 - 4: Merged signal (red and green). Green - ab193379 observed at 30 kDa. Red - loading control, Anti-GAPDH antibody [EPR16891] - Loading Control ab181602, observed at 37 kDa.

  ab193379 was shown to specifically recognize CDKN1B in wild-type HAP1 cells as well as additional cross-reactive bands. No bands were observed when CDKN1B knockout samples were used. Wild-type and CDKN1B knockout samples were subjected to SDS-PAGE. ab193379 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 μg/mL and 1/10,000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-p27 KIP 1 antibody [SX53G8] (ab193379)

  Predicted band size: 22 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [SX53G8] (ab193379)

  Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human colon tissue labeling p27 KIP 1 with ab193379 at 1 μg/mL.

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  Western blot - Anti-p27 KIP 1 antibody [SX53G8] (ab193379)

  Western blot: Anti-CDKN1B antibody [SX53G8] (ab193379) staining at 1 ug/ml, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (Anti-alpha Tubulin antibody [EP1332Y] - Loading Control ab52866) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab193379 was shown to bind specifically to CDKN1B. A band was observed at 30 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CDKN1B knockout cell line. To generate this image, wild-type and CDKN1B knockout HCT 116 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-p27 KIP 1 antibody [SX53G8] (ab193379) at 1 µg/mL

  Lane 1: Wild-type HCT 116 cell lysate at 20 µg

  Lane 2: CDKN1B knockout HCT 116 cell lysate at 20 µg

  Lane 3: Wild-type HAP1 cell lysate at 20 µg

  Lane 4: CDKN1B knockout HAP1 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: 30 kDa