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KO Validation of CDKN2A in 293 Cells. Loading: 10 µg of 293 WT cell lysates or CDKN2A KO cell lysates. Antibodies: CDKN2A, 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. |
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Immunohistochemistry Validation of CDKN2A in Human Colon Tissue. Immunohistochemical analysis of paraffin-embedded Human Colon Tissue using anti- CDKN2A 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 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. |
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Immunohistochemistry Validation of CDKN2A in Rat Colon Tissue. Immunohistochemical analysis of paraffin-embedded Rat Colon Tissue using anti- CDKN2A 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 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. |
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Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: CDKN2A orb1239282 (4 µg/mL), CDKN2A orb1255478 (4 µg/mL), and beta-actin (1 µg/mL), overnight incubation at 4C in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
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Recombinant Protein Test. Loading: 15 µg of lysates per lane. Antibodies: CDKN2A orb1239282 (2 µg/mL), overnight incubation at 4C 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 recombinant protein (arrow: monomer and dimer). |
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Western Blot Validation in Human Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: CDKN2A, orb1239282 (2 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
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Western Blot Validation in Human Normal and Cancer Tissue. Loading: 15 µg of lysates per lane. Antibodies CDKN2A orb1239282 (2 µg/mL), overnight incubation at 4C 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. |
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Western Blot Validation in Mouse Colon Tissue. Loading: 15 µg of lysates per lane. Antibodies: CDKN2A 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. |
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Immunofluorescence Validation of CDKN2A in Human A431 Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed A431 cells labeling CDKN2A with orb1239282 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue). |
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Immunofluorescence Validation of CDKN2A in Human Colon Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed human colon tissue labeling CDKN2A with orb1239282 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue). |
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Immunofluorescence Validation of CDKN2A in Rat Colon Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed Rat Colon Tissue labeling CDKN2A with orb1239282 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue). |