CDKN2A/p16INK4a Antibody

Immunohistochemistry Validation of CDKN2A/p16INK4a in Rat Colon Tissue. Immunohistochemical analysis of paraffin-embedded Rat Colon Tissue using anti- CDKN2A/p16INK4a antibody (orb1239282) at 5 µg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunohistochemistry Validation of CDKN2A/p16INK4a in Human Colon Tissue. Immunohistochemical analysis of paraffin-embedded Human Colon Tissue using anti- CDKN2A/p16INK4a antibody (orb1239282) at 10 µg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunofluorescence Validation of CDKN2A/p16INK4a in Rat Colon Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed Rat Colon Tissue labeling CDKN2A/p16INK4a with orb1239282 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).

Immunofluorescence Validation of CDKN2A/p16INK4a in Human Colon Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed human colon tissue labeling CDKN2A/p16INK4a with orb1239282 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Immunofluorescence Validation of CDKN2A/p16INK4a in Human A431 Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed A431 cells labeling CDKN2A/p16INK4a with orb1239282 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Western Blot Validation in Mouse Colon Tissue. Loading: 15 µg of lysates per lane. Antibodies: CDKN2A/p16INK4a orb1239282 (A: 1 µg/mL, B: 2 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Western Blot Validation in Human Normal and Cancer Tissue. Loading: 15 µg of lysates per lane. Antibodies CDKN2A/p16INK4a orb1239282 (2 µg/mL), overnight incubation at 4°C in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: Human breast, Lane 2: Human lung tumor, Lane 3: Human colon cancer.

Western Blot Validation in Human Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: CDKN2A/p16INK4a, orb1239282 (2 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Recombinant Protein Test. Loading: 15 µg of lysates per lane. Antibodies: CDKN2A/p16INK4a orb1239282 (2 µg/mL), overnight incubation at 4°C in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: Human prostate (benign hyperplasia) Lane 2: Human CDKN2A/p16INK4a recombinant protein (arrow: monomer and dimer).

Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: CDKN2A/p16INK4a orb1239282 (4 µg/mL), CDKN2A/p16INK4a orb1255478 (4 µg/mL), and beta-actin (1 µg/mL), overnight incubation at 4°C in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

KO Validation of CCDKN2A/p16INK4a in 293 Cells. Loading: 10 µg of 293 WT cell lysates or CDKN2A/p16INK4a KO cell lysates. Antibodies: CDKN2A/p16INK4a, orb1239282 (2 µg/mL) and beta-actin, orb1240312 (1 µg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.