![Western blot - p16 INK4a antibody [2D9A12] (ab54210) image](#ab54210_1.jpg)
![Immunohistochemistry (Paraffin-embedded sections) - CDKN2A/p16INK4a antibody [2D9A12] (ab54210) image](#ab54210_2.jpg)
![Immunohistochemistry (Paraffin-embedded sections) - CDKN2A/p16INK4a antibody [2D9A12] (ab54210) image](#ab54210_3.jpg)
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Products:Cell Biology >> Cell Cycle >> Cell Cycle Inhibitors >> Ink4
Overview
Product name
Anti-CDKN2A/p16INK4a antibody [2D9A12]
See all CDKN2A/p16INK4a products (11) ...
Description
Mouse monoclonal [2D9A12] to CDKN2A/p16INK4a
Tested applications
Flow Cyt, WB, ELISA, IHC-Pmore details
Cross reactivity
Reacts with
Rat, Human
Immunogen
Ni-NTA purified recombinant P16 expressed in E. Coli strain BL21 (DE3).
Positive control
Rat liver tissue; Human brain tissue; Human brain tumor tissue.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer
Preservative: 0.03% Sodium Azide
Constituents: Ascites
Concentration
Concentration information loading...
Purity
Ascites
Clonality
Monoclonal
Clone number
2D9A12
Isotype
IgG2b
Research areas
Cancer >> Oncoproteins/suppressors >> Tumor suppressors >> INK4a/Arf
Cancer >> Cell cycle >> Cell cycle inhibitors >> Ink4
Epigenetics and Nuclear Signaling >> Cell cycle >> Cell Cycle Inhibitors >> Ink4
Cell Biology >> Cell Cycle >> Cell Cycle Inhibitors >> Ink4
Applications
Show applications key
Our Abpromise guarantee covers the use of ab54210 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Flow Cyt: Use 1µg for 106 cells.
WB: 1/500 - 1/2000.(Tested using a recombinant protein expressed in E. coli. We have no data on the detection of the endogenous protein and would welcome feedback from anyone that has performed a WB with ab54210 using cell lysates.)
ELISA
ELISA: 1/10000ELISA: 1/10000
IHC-P: 1/200 - 1/1000.Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.(Recommended antigen retrieval: Place slides in a special glass slide holder and fill in the rack with slides to ensure even heating. Rinse rack in 1L of Citrate buffer in ordinary pressure cooker,(0.01M,pH6.0) to immerse all slides. Cover the cap and heat for 15-20 minutes on electric cooker until water boils. Begin timing, only 2 minutes in high-pressure Remove the pressure cooker, Allow it to cool(Cool the slides at room temperature) Rinse the slides in PBS for 5 minutes)
Target
Relevance
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. In spite of the structural and some functional differences, all the proteins encoded by the CDKN2A gene are involved in cell cycle G1 control.
Cellular localization
Cytoplasmic and Nuclear
Form
There are 4 isoforms produced by alternative splicing. Isoform 1 also known as: p16INK4a; Isoform 3 also known as: p12; Isoform 4 also known as: p14ARF; p19ARF; ARF.
Alternative names
- INK4 antibody
- P16INK4 antibody
- ARF antibody
see all
Database links
- Entrez Gene: 1029 Human
- Entrez Gene: 25163 Rat
- Omim: 600160 Human
- SwissProt: P42771 Human
- SwissProt: Q9R0Z3 Rat
- Unigene: 512599 Human
- Unigene: 48717 Rat
Anti-CDKN2A/p16INK4a antibody [2D9A12] images:
Western blot - p16 INK4a antibody [2D9A12] (ab54210)
![Western blot - p16 INK4a antibody [2D9A12] (ab54210)](/ps/datasheet/Images/54/ab54210/ab54210_1.jpg)
All lanes : Anti-CDKN2A/p16INK4a antibody [2D9A12] (ab54210) at 1/500 dilution
Lane 1 : Molecular weight marker
Lane 2 : Truncated p16 recombinant protein
Observed band size : 40 kDa (why is the actual band size different from the predicted?)
Immunohistochemistry (Paraffin-embedded sections) - CDKN2A/p16INK4a antibody [2D9A12] (ab54210)
![Immunohistochemistry (Paraffin-embedded sections) - CDKN2A/p16INK4a antibody [2D9A12] (ab54210)](/ps/datasheet/Images/54/ab54210/ab54210_2.jpg)
Ab54210 (1:500) staining p16 INK4a in rat liver tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (by DAB staining) is shown.
Immunohistochemistry (Paraffin-embedded sections) - CDKN2A/p16INK4a antibody [2D9A12] (ab54210)
![Immunohistochemistry (Paraffin-embedded sections) - CDKN2A/p16INK4a antibody [2D9A12] (ab54210)](/ps/datasheet/Images/54/ab54210/ab54210_3.jpg)
Ab54210 (1:500) staining p16 INK4a in human brain tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (by DAB staining) is shown.
Immunohistochemistry (Paraffin-embedded sections) - CDKN2A/p16INK4a antibody [2D9A12] (ab54210)
![Immunohistochemistry (Paraffin-embedded sections) - CDKN2A/p16INK4a antibody [2D9A12] (ab54210)](/ps/datasheet/Images/54/ab54210/ab54210_4.jpg)
Ab54210 (1:500) staining p16 INK4a in human brain tumor tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (by DAB staining) is shown.
Flow Cytometry-CDKN2A/p16INK4a antibody [2D9A12](ab54210)
](/ps/datasheet/images/54/ab54210/CDKN2Ap16INK4a-Primary-antibodies-ab54210-2.jpg)
Overlay histogram showing HeLa cells stained with ab54210 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab54210, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.
References for Anti-CDKN2A/p16INK4a antibody [2D9A12] (ab54210)
This product has been referenced in:
- Royds JAet al. The CDKN2A G500 Allele Is More Frequent in GBM Patients with No Defined Telomere Maintenance Mechanism Tumors and Is Associated with Poorer Survival. PLoS One 6:e26737 (2011). IHC-P; Human.Read more (PubMed: 22046342) »
- Almeida MQet al. Mouse Prkar1a haploinsufficiency leads to an increase in tumors in the Trp53+/- or Rb1+/- backgrounds and chemically induced skin papillomas by dysregulation of the cell cycle and Wnt signaling. Hum Mol Genet 19:1387-98 (2010). IHC-P; Mouse.Read more (PubMed: 20080939) »
See all 3 publications for this product
Publishing research using ab54210? Please let us know so that we can cite the reference in this datasheet
![CDKN2A/p16INK4a antibody [2D9A12] for IHC-P in Human (54210)](/ps/Reviews/Images/ab12857_72508.jpg)
![CDKN2A/p16INK4a antibody [2D9A12] for IHC-P in Human (54210)](/ps/datasheet/images/54/ab54210/CDKN2Ap16INK4a-Primary-antibodies-ab54210-1.gif)
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"