CDKN2A/p16INK4a Rabbit Polyclonal Antibody

Western blot analysis of CDKN2A/p16INK4a on different lysates with Rabbit anti-CDKN2A/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: 17 kDa
Observed band size: 16 kDa

Exposure time: 1 minute;

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.

Immunocytochemistry analysis of HeLa cells labeling CDKN2A/p16INK4a with Rabbit anti-CDKN2A/p16INK4a antibody (ER1803-53) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-CDKN2A/p16INK4a antibody (ER1803-53) at 1/100 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 (M1305-2, 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.

ICC staining CDKN2A/p16INK4a in 293T cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

ICC staining CDKN2A/p16INK4a in SiHa cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

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 (pH6) for 20 mins.

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 (pH6) for 20 mins.

Flow cytometric analysis of 293T cells with CDKN2A/p16INK4a antibody at 1/100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti-rabbit IgG was used as the secondary antibody.

☑ Knockdown (KD)

Western blot analysis of CDKN2A/p16INK4a on different lysates with Rabbit anti-CDKN2A/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.