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<strong>Figure 1 KO Validation of CDKN2A in 293 Cells</strong><br> Loading: 10 μg of 293 WT cell lysates or CDKN2A KO cell lysates. Antibodies: CDKN2A, 8975 (2 μg/mL) and beta-actin, 3779 (1 μg/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2. Immunofluorescence Validation of CDKN2A In HeLa Cells </strong><br> Immunofluorescent analysis of methanol-fixed HeLa cells labeling CDKN2A with 8975 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and Hoechst staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.
<strong>Figure 3 Independent Antibody Validation (IAV) via Protein Expression Profile in Human Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: CDKN2A, 4211 (2 μg/mL), CDKN2A, 8975 (2 μg/mL) and beta-actin, 3779 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Western Blot Validation in Human Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: CDKN2A, 8975 (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 5 Immunohistochemistry Validation of CDKN2A in Human Colon Tissue</strong><br> Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-CDKN2A antibody (8975) 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.
Flow Cytometry Validation of CDKN2A in HeLa Cells 1X10^6/mL HeLa cells were intracellularly stained with 2 μg/mL of CDKN2A antibody (red) or Rabbit IgG isotype control (black) at RT for 30 min, followed by PSFlo 488 conjugated goat anti-rabbit IgG secondary antibody staining (1:1000) at RT for 30 min. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100 in PBS.
Figure 1 KO Validation of CDKN2A in 293 Cells
Loading: 10 μg of 293 WT cell lysates or CDKN2A KO cell lysates. Antibodies: CDKN2A, 8975 (2 μg/mL) and beta-actin, 3779 (1 μg/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
IAV
KO
New
Trial Size

CDKN2A/p16INK4a Antibody

CATALOG NUMBER: 8975

Clonality:
Polyclonal
Tested Applications:
ELISA, FC, IHC-P, WB, ICC/IF
Host Species:
Rabbit
Species Reactivity:
Human
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human
Immunogen:
Anti-CDKN2A antibody (8975) was raised against a peptide corresponding to 17 amino acids near the carboxy terminus of human CDKN2A.
The immunogen is located within the last 50 amino acids of CDKN2A.
Conjugate:
Unconjugated
Tested Applications:
ELISA, FC, IHC-P, WB, ICC/IF
Application Note:
WB: 1 - 2 μg/mL; IHC: 5 μg/mL; IF: 20 μg/mL; FC: 2 μg/mL

Antibody validated: Western Blot in human samples; Immunohistochemistry and Immunofluorescence in human samples; Flow Cytometry in human samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1201 - HeLa Cell Lysate
Positive Control 2:
Cat. No. 1211 - HepG2 Cell Lysate
Predicted Molecular Weight:
Predicted: 17kD
Observed: 17kD
Validation:

KO Validation (Figure 1): Anti-CDKN2A antibodies (8975) specificity was further verified by CDKN2A specific knockout. CDKN2A signal was not detected in CDKN2A knockout 293 cells as compared to that in control wild type cells.

Independent Antibody Validation in Cell lines (Figure 2) shows similar CDKN2A expression profile in human cell lines detected by two independent anti-CDKN2A antibodies that recognize different epitopes, 4211 against N-terminus domain and 8975 against the C-terminus domain.  CDKN2A proteins are detected only in 293, HeLa and HepG2 cell lines by the two independent antibodies. 

Isoforms:
Human CDKN2A has 6 isoforms, including isoform 1 (156aa, 17kD), isoform 2 (105aa, 11kD), isoform 3 (116aa, 12kD), isoform 5 (167aa, 18kD), isoform tumor suppressor ARF (132aa, 14kD) and isoform smARF (85aa, 9kD). Mouse CDKN2A has 4 isoforms, including isoform 1 (168aa, 18kD), isoform 2 (125aa, 14kD), isoform tumor suppressor ARF (169aa, 19kD) and isoform smARF (125aa, 14kD). Rat CDKN2A has two isoforms, including isoform 1 (159aa, 17kD) and isoform ARF (160aa, 17kD). 8975 can detect only human isoforms 1, 2 and 5, but cannot detect mouse and rat.
Purification:
CDKN2A Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
CDKN2A Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
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.
Ncbi Official Symbol:
CDKN2A
Additional Names:
CDKN2A Antibody: P16, INK4, MTS1, CDK4I, CDKN2, INK4A, MTS-1, P16INK4, P16INK4A, P16-INK4A, Cyclin-dependent kinase 4 inhibitor A
Protein Accession Number:
NP_000068
Protein Gi Number:
4502749
Ncbi Gene Id Number:
1029
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
CDKN2A Antibody: The CDKN2A locus gives rise to 2 distinct transcripts from different promoters. The transcripts have been designated p16(INK4A) and p14(ARF). This chromosomal region undergoes a number of inversions, translocations, heterozygous deletions, and homozygous deletions in a variety of malignant cell lines including those from glioma, non-small cell lung cancer, leukemia, and melanoma. Deletion of the region containing CDKN2A is found in more than half of all melanoma cell lines. Conversely, transfection of CDKN2A suppressed the growth of two independent mesothelioma cell lines, suggesting that inactivation of the CDKN2 gene is an essential step in the etiology of malignant mesotheliomas. CDKN2A induces a G1 cell cycle arrest by inhibiting the phosphorylation of the Rb protein by the cyclin-dependent kinases CDK4 and CDK6. CDKN2A is expressed as at least three distinct isoforms.
Background Reference 1:
Stone et al. Cancer Res. 1995; 55:2988-94.
Background Reference 2:
Kamb et al. Nature Genet. 1994; 8:22-6.
Background Reference 3:
Kratzke et al. J. Nat. Cancer Inst. 1995; 87:1870-5.
Background Reference 4:
Stott et al. EMBO J. 1998; 17:5001-14.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

8975

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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