Cervical lymph node metastasis of oropharyngeal HPV-associated squamous cell
((A), high grade squamous intraepithelial lesion of the uterine cervix (in contrast with normal endocervical glands))
Rabbit anti-Human p16 Monoclonal Antibody | anti-p16 antibody
Anti - p16
1. Deparaffinize the section in 3 changes of xylene, 10 minutes each.
2. Wash the section in 96%, 80% and 70% ethyl alcohol for 10 minutes each.
3. Rinse in distilled water, 2 x 5 minutes.
4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes.
5. Wash in distilled water, 2 x 5 minutes.
6. For antigen retrieval: Immerse the slide in Tris-EDTA buffer*, pH 9.0 and incubate at 95-97ºC in water bath for 25 minutes.
7. Remove the staining to room temperature and let the slide to cool (in Tris-EDTA buffer, pH 9.0) for 15 minutes.
8. Rinse in distilled water, 2 x 5 minutes.
9. Wash in PBS (phosphate buffer saline, pH 7.0-7.5) supplemented with 0.05% of Tween-20 (buffer A), 2 x 5 minutes.
10. CONCENTRATED:
Incubate the section with primary antibody at the dilution 1:100 for 1 hour in the closed wet chamber.
READY TO USE (RTU):
Incubate the section with primary antibody (ready to use) for 1 hour in a closed wet chamber.
11. Wash 3 x 5 minutes with buffer A.
12. Apply the secondary antibody (the protocol depends on the supplier), and proceed to standard immunohistochemistry protocol (HRP - Peroxide - DAB). Micropolymer-HRP detection kit rabbit/mouse dual of MBS is suggested
13. Wash 3 x 5 minutes with buffer A.
14. Apply the chromogen (DAB), 1 - 3 minutes.
15. Wash in water, 2 x 5 minutes.
16. Stain in hematoxylin for 5 minutes.
17. Wash in distilled water, 3 x 2 minutes.
18. Mount the slide for observation.
Tris - EDTA Buffer (10mM Tris Base, 1mM EDTA solution, pH 9.0):
Tris ----- ------1.21 g; EDTA --------- 0. 37 g; Distilled water ----------- 1000 ml Mix to dissolve in 700 ml of distilled water. Adjust pH to 9.0 with 1M HCl and mix well. Adjust the final volume to 1 liter with distilled water.
Store this solution at room tem perature for 3 months or at +4º C for longer storage.
1. Drying (Enter).
2. Heating (72°C), incubation 4 min; drying.
3. Deparafinization (Enter).
4. Heating (72°C) with the medium temperatures.
5. Cell conditioning (Enter).
6. ULTRA conditioner #1 (Enter).
7. Heating glass (95°C), incubation 8 min (Cell conditioner #1).
8. ULTRA CC1 solution application –20 min (Enter).
9. ULTRA CC1 solution application –36 min (Enter).
10. Titration (Enter).
11. Hand Apply –primary antibody. Incubation 36 min.
12. Nuclear stain (Enter).
13. Hematoxylin application – one drop (Nuclear stain). Cover and incubate 4 min.
14. After nuclear stain (Enter).
15. Bluing reagent application, one drop. After nuclear stain, cover and incubate 4 min.
2. Centrifuge the vial before use.
3. Intended for Research use only!
4. Do not use after expiration date stamped on vial label.
5. Avoid contamination of the reagent.
6. Any discrepancies in the recommended procedures stated in the working protocol may affect the final results.
7. The reagent contains sodium azide (NaN3) which is highly toxic in higher concentrations. The concentration in the reagent (0.05%) is not considered as hazardous.
8. Disposal of waste material must be conducted in accordance with local regulations.
9. Wear appropriate Personal Protective Equipment to avoid contact with eyes and skin.
Cervical lymph node metastasis of oropharyngeal HPV-associated squamous cell
((A), high grade squamous intraepithelial lesion of the uterine cervix (in contrast with normal endocervical glands))
Cervical lymph node metastasis of oropharyngeal HPV-associated squamous cell
((A), high grade squamous intraepithelial lesion of the uterine cervix (in contrast with normal endocervical glands))
Testing Data #2
((B), high grade squamous intraepithelial lesion of the uterine cervix (in contrast with non-dysplastic metaplastic squamous epithelium))
Testing Data #2
((B), high grade squamous intraepithelial lesion of the uterine cervix (in contrast with non-dysplastic metaplastic squamous epithelium))
Testing Data #3
((C), adenocarcinoma in situ of the uterine cervix)
Testing Data #3
((C), adenocarcinoma in situ of the uterine cervix)
Testing Data #4
((D), stained with anti-p16 monospecific clonal antibody. All show strong and specific positive immunostaining of dysplastic and neoplastic epithelium, with no reactivity in normal epithelial and stromal structures. Formalin fixed, paraffin embedded human tissues (4 um sections) stained according to MyBioSource.)
Testing Data #4
((D), stained with anti-p16 monospecific clonal antibody. All show strong and specific positive immunostaining of dysplastic and neoplastic epithelium, with no reactivity in normal epithelial and stromal structures. Formalin fixed, paraffin embedded human tissues (4 um sections) stained according to MyBioSource.)
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
Function: Acts as a negative regulator of the proliferation of normal cells by interacting strongly with CDK4 and CDK6. This inhibits their ability to interact with cyclins D and to phosphorylate the retinoblastoma protein. Ref.11 Ref.13
Subunit structure: Heterodimer with CDK4 or CDK6. Predominant p16 complexes contained CDK6. Interacts (isoforms 1,2 and 4) with CDK4 (both 'T-172'-phosphorylated and non-phosphorylated forms); the interaction inhibits cyclin D-CDK4 kinase activity. Interacts with ISCO2. Ref.13 Ref.14
Subcellular location: Cytoplasm. Nucleus Ref.14.
Tissue specificity: Widely expressed but not detected in brain or skeletal muscle. Isoform 3 is pancreas-specific. Ref.2
Post-translational modification: Phosphorylation seems to increase interaction with CDK4.
Involvement in disease: The association between cutaneous and uveal melanomas in some families suggests that mutations in CDKN2A may account for a proportion of uveal melanomas. However, CDKN2A mutations are rarely found in uveal melanoma patients.Melanoma, cutaneous malignant 2 (CMM2) [MIM:155601]: A malignant neoplasm of melanocytes, arising de novo or from a pre-existing benign nevus, which occurs most often in the skin but also may involve other sites.Note: Disease susceptibility is associated with variations affecting the gene represented in this entry. Ref.24 Ref.27 Ref.29 Ref.30 Ref.31 Ref.32 Ref.34 Ref.36 Ref.38 Ref.39 Ref.41 Ref.42Familial atypical multiple mole melanoma-pancreatic carcinoma syndrome (FAMMMPC) [MIM:606719]: An inherited cancer predisposition syndrome characterized by an increased risk of developing malignant melanoma and/or pancreatic cancer. Mutation carriers within families may develop either or both types of cancer.Note: The disease is caused by mutations affecting the gene represented in this entry.Li-Fraumeni syndrome (LFS) [MIM:151623]: Autosomal dominant familial cancer syndrome that in its classic form is defined by the existence of a proband affected by a sarcoma before 45 years with a first degree relative affected by any tumor before 45 years and another first degree relative with any tumor before 45 years or a sarcoma at any age. Other clinical definitions for LFS have been proposed (PubMed:8118819 and PubMed:8718514) and called Li-Fraumeni like syndrome (LFL). In these families affected relatives develop a diverse set of malignancies at unusually early ages. Four types of cancers account for 80% of tumors occurring in TP53 germline mutation carriers: breast cancers, soft tissue and bone sarcomas, brain tumors (astrocytomas) and adrenocortical carcinomas. Less frequent tumors include choroid plexus carcinoma or papilloma before the age of 15, rhabdomyosarcoma before the age of 5, leukemia, Wilms tumor, malignant phyllodes tumor, colorectal and gastric cancers.Note: The disease is caused by mutations affecting the gene represented in this entry. Ref.35Melanoma-astrocytoma syndrome (MASTS) [MIM:155755]: Characterized by a dual predisposition to melanoma and neural system tumors, commonly astrocytoma.Note: The disease is caused by mutations affecting the gene represented in this entry. Ref.37
Sequence similarities: Belongs to the CDKN2 cyclin-dependent kinase inhibitor family.Contains 4 ANK repeats.
Sequence caution: The sequence AAB60645.1 differs from that shown. Reason: Erroneous initiation.
Research Articles on p16
Similar Products
Product Notes
The p16 cdkn2a (Catalog #AAA684198) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti - p16 reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's p16 can be used in a range of immunoassay formats including, but not limited to, Immunohistochemistry (IHC) - Formalin/Paraffin. IHC-P dilution 1:100. Researchers should empirically determine the suitability of the p16 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 "p16, Monoclonal 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
If you are ready to order, navigate to Shopping Cart and get ready to checkout.
