CDKN2A Antibody

• Catalog Number: orb1239282
• Category: Antibodies
• Description: CDKN2A Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC-P, WB
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: Anti-CDKN2A antibody (orb1239282) was raised against a peptide corresponding to 18 amino acids near the amino terminus of human CDKN2A. The immunogen is located within the first 50 amino acids of CDKN2A.
• Concentration: 1 mg/mL
• Dilution range: WB: 1 - 4 μg/mL; IHC-P: 5 - 10 μg/mL; IF: 20 μg/mL.Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry in human and rat samples; Immunofluorescence in human and rat samples. All other applications and species not yet tested.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 17 kDa Observed: 17 kDa
• Target: CDKN2A
• UniProt ID: P42771
• NCBI: NP_000068
• Storage: CDKN2A antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
• Buffer/Preservatives: CDKN2A Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: CDKN2A Antibody: ARF, MLM, P14, P16, P19, CMM2, IN
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

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.

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 4°C 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 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 recombinant protein (arrow: monomer and dimer).

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.

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 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 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.

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).

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).

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).

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 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 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 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.