Rabbit CDKN2A / p16INK4a Polyclonal Antibody | anti-CDKN2A antibody
Rabbit Polyclonal (IgG) to Human CDKN2A / p16INK4a
IHC-Paraffin (5 ug/ml)
Western Blot (1 - 2 ug/ml)
Optimal dilution to be determined by the researcher.
Usage Summary: CDKN2A antibody can be used for detection of DKN2A by Western Blot at 1-2 ug/mL. Antibody can also be used for immunohistochemistry starting at 5ug/mL. For immunofluorescence start at 20 ug/mL. Antibody validated: Western Blot in mouse samples; Immunohistochemistry in human and rat samples. All other applications and species are not yet tested. Western Blot: Predicted: 17 kDa Observed: 20 kDa
Tissue Preparation:
Formalin fixation and embedding in paraffin wax.
Tissue Sectioning:
Make 4-um sections and place on pre-cleaned and charged microscope slides. Heat in a tissue-dryingoven for 45 minutes at 60°C.
Deparaffinization:
Wash dry slides in 3 changes of xylene - 5 minutes each @ RT
Rehydration:
Wash slides in 3 changes of 100% alcohol - 3 minutes each @ RT
Wash slides in 2 changes of 95% alcohol - 3 minutes each @ RT
Wash slides in 1 change of 80% alcohol - 3 minutes @ RT
Rinse slides in gentle running distilled water - 5 minutes @ RT
Antigen retrieval:
Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes
Remove from heat and let stand at room temperature in buffer - 20 minutes
Rinse in 1X TBS with Tween (TBST) -1 minute @ RT
Immunostaining:
(Do not allow tissues to dry at any time during the staining procedure)
Apply a universal protein block - 20 minutes @ RT
Drain protein block from slides, apply diluted primary antibody - 45 minutes @ RT
Rinse slides in 1 X TBST - 1 minute @ RT
Apply a biotinylated secondary antibody appropriate for the primary antibody - 30 minutes @ RT
Rinse slides in 1X TBST -1 minute @ RT
Apply alkaline phosphatase streptavidin - 30 minutes @ RT
Rinse slides in 1X TBST -1 minute @ RT
Apply alkaline phosphatase chromogen substrate - 30 minutes @ RT
Wash slides in distilled water - 1 minute @ RT
Dehydrate:
(This method should only be used if the chromogen substrate is alcohol insoluble (e.g. Vector Red, DAB)
Wash slides in 2 changes of 80% alcohol - 1 minute each @ RT
Wash slides in 2 changes of 95% alcohol - 1 minute each @ RT
Wash slides in 3 changes of 100% alcohol - 1 minute each @ RT
Wash slides in 3 changes of xylene - 1 minute each @ RT
Apply coverslip
NCBI and Uniprot Product Information
NCBI Description
This gene generates several transcript variants which differ in their first exons. At least three alternatively spliced variants encoding distinct proteins have been reported, two of which encode structurally related isoforms known to function as inhibitors of CDK4 kinase. The remaining transcript includes an alternate first exon located 20 Kb upstream of the remainder of the gene; this transcript contains an alternate open reading frame (ARF) that specifies a protein which is structurally unrelated to the products of the other variants. This ARF product functions as a stabilizer of the tumor suppressor protein p53 as it can interact with, and sequester, the E3 ubiquitin-protein ligase MDM2, a protein responsible for the degradation of p53. In spite of the structural and functional differences, the CDK inhibitor isoforms and the ARF product encoded by this gene, through the regulatory roles of CDK4 and p53 in cell cycle G1 progression, share a common functionality in cell cycle G1 control. This gene is frequently mutated or deleted in a wide variety of tumors, and is known to be an important tumor suppressor gene. [provided by RefSeq, Sep 2012]
Uniprot Description
p16-INK4A: a cell-cycle regulatory protein that interacts with CDK4 and CDK6, inhibiting their ability to interact with cyclins D. Inhibits the phosphorylation of the retinoblastoma protein by CDK4 or CDK6, and entry into the S phase of the cell cycle. The p16INK4A and p14ARF proteins are encoded by CDKN2A, a known tumour suppressor gene in multiple cancers. CDKN2A is inactivated in 72% of cases of lung squamous cell carcinoma: 21% by epigenetic silencing by methylation, 18% inactivating mutation, 4% by exon 1b skipping, and 29% by homozygous deletion. Defects in CDKN2A are the cause of familial atypical multiple mole melanoma-pancreatic carcinoma syndrome, Li-Fraumeni syndrome, and the melanoma-astrocytoma syndrome. The melanoma-astrocytoma syndrome is characterized by a dual predisposition to melanoma and neural system tumors, commonly astrocytoma. Four alternatively spliced p16 isoforms have been reported. Two alternatively spliced isoforms of ARF have been reported.
Protein type: Cell cycle regulation; Tumor suppressor; Nucleolus
Chromosomal Location of Human Ortholog: 9p21
Cellular Component: nucleoplasm; nuclear body; protein complex; mitochondrion; cytoplasm; nucleolus; nucleus; cytosol
Molecular Function: cyclin-dependent protein kinase inhibitor activity; NF-kappaB binding; protein binding; DNA binding; p53 binding; ubiquitin-protein ligase inhibitor activity; protein kinase binding; transcription factor binding
Biological Process: protein polyubiquitination; positive regulation of apoptosis; positive regulation of transcription, DNA-dependent; regulation of protein stability; negative regulation of B cell proliferation; regulation of protein export from nucleus; negative regulation of cell proliferation; apoptotic mitochondrial changes; regulation of G2/M transition of mitotic cell cycle; somatic stem cell division; cell cycle arrest; caspase activation; negative regulation of immature T cell proliferation in the thymus; protein stabilization; protein destabilization; transcription, DNA-dependent; negative regulation of cyclin-dependent protein kinase activity; negative regulation of cell-matrix adhesion; positive regulation of protein sumoylation; inhibition of NF-kappaB transcription factor; Ras protein signal transduction; negative regulation of ubiquitin-protein ligase activity; negative regulation of protein kinase activity; negative regulation of phosphorylation; positive regulation of transcription from RNA polymerase II promoter; mitotic cell cycle; positive regulation of DNA damage response, signal transduction by p53 class mediator; negative regulation of cell growth; negative regulation of transcription, DNA-dependent; rRNA processing; G1/S transition of mitotic cell cycle
Disease: Melanoma-astrocytoma Syndrome; Melanoma, Cutaneous Malignant, Susceptibility To, 2; Melanoma-pancreatic Cancer Syndrome
Research Articles on CDKN2A
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Product Notes
The CDKN2A cdkn2a (Catalog #AAA241263) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's CDKN2A / p16INK4a can be used in a range of immunoassay formats including, but not limited to, Immunohistochemistry - Paraffin (IHC-P), Western Blot (WB), Immunofluorescence (IF), ELISA (EIA). Usage: IHC-Paraffin (5 ug/ml) Western Blot (1 - 2 ug/ml) Optimal dilution to be determined by the researcher. Usage Summary: CDKN2A antibody can be used for detection of DKN2A by Western Blot at 1-2 ug/mL. Antibody can also be used for immunohistochemistry starting at 5ug/mL. For immunofluorescence start at 20 ug/mL. Antibody validated: Western Blot in mouse samples; Immunohistochemistry in human and rat samples. All other applications and species are not yet tested. Western Blot: Predicted: 17 kDa Observed: 20 kDa. Researchers should empirically determine the suitability of the CDKN2A cdkn2a for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "CDKN2A / p16INK4a, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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