p16INK4a Rabbit Polyclonal Antibody

Western blot analysis of p16INK4a on different lysates with Rabbit anti-p16INK4a antibody (ER1803-53) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HEK-293 cell lysate
Lane 3: Saos-2 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 16 kDa
Observed band size: 16 kDa

Exposure time: 1 minute; ECL: K1801;

15% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER1803-53) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

☑ Knockdown (KD)

Western blot analysis of p16INK4a on different lysates with Rabbit anti-p16INK4a antibody (ER1803-53) at 1/1,000 dilution.

Lane 1: HeLa-si NT cell lysate
Lane 2: HeLa-si p16INK4a cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 16 kDa
Observed band size: 16 kDa

Exposure time: 9 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER1803-53) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of HeLa cells labeling p16INK4a with Rabbit anti-p16INK4a antibody (ER1803-53) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-p16INK4a antibody (ER1803-53) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-CDKN2A/p16INK4a antibody. Counter stained with hematoxylin. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes.

Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue using anti-CDKN2A/p16INK4a antibody. Counter stained with hematoxylin. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes.

Flow cytometric analysis of HeLa cells labeling p16INK4a.

Cells were fixed and permeabilized. Then stained with the primary antibody (ER1803-53, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).