The MTS1/CDKN2/p16 gene encoding the p16INK4a tumor-suppressor protein is usually inactivated by homozygous deletion or hypermethylation of the promoter in a variety of human malignancies. In choose tumor varieties, together with pancreatic adenocarcinomas, intragenic mutations are present in a big share of instances.
The immunoreactivity of mutant p16 proteins has not been comprehensively studied. Furthermore, the immunohistochemical properties of commercially obtainable antibodies haven’t been described intimately. We studied 35 pancreatic adenocarcinomas with a molecularly outlined p16 standing (16 homozygous deletions, Three hypermethylated instances, and 16 tumors with an intragenic mutation in a single allele related to lack of the second allele).
As well as, we studied 9 cell traces (three homozygous deletions, three hypermethylated traces, and three intragenic mutations). Paraffin sections of the tumors and cell blocks had been reacted with 4 totally different anti-p16 antibodies: polyclonal and monoclonal (clone G175-405) antibodies from PharMingen, monoclonal antibody DCS-50 from Oncogene Science, and monoclonal antibody ZJ11 from Neo-Markers.
Optimum staining situations had been established for every antibody. The pancreatic carcinomas with homozygous p16 deletions had been largely devoid of nuclear staining (admixed nonneoplastic cells served as inside constructive controls); just one adenocarcinoma every reacted with DCS-50 and the polyclonal antibody, and 5 had been constructive with ZJ11, suggesting that nonspecific nuclear staining can happen below sure situations.
Antibody DCS-50 produced nuclear staining in all three hypermethylated carcinomas, whereas G175-405 stained none of them. Three of the 4 antibodies produced nuclear immunoreactivity in 7 to 14 of the 16 carcinomas carrying p16 mutations; G175-405 confirmed solely weak reactivity in a single case.
Cytoplasmic staining was current in all carcinomas and cell traces and with all antibodies and subsequently can’t be thought of particular; it was strongest with G175-405. Thus, we discovered antibody G175-405 to be probably the most particular, and monoclonals DCS-50 and ZJ11 the least particular for wild-type p16. Nevertheless, the previous tends to offer stronger cytoplasmic background staining. For tumor varieties wherein p16 mutations are unusual, the PharMingen polyclonal antibody could also be an appropriate various.
The specificity and patterns of staining in human cells and tissues of p16INK4a antibodies show variant antigen binding.
The validity of the identification and classification of human most cancers utilizing antibodies to detect biomarker proteins relies upon upon antibody specificity. Antibodies that bind to the tumour-suppressor protein p16INK4a are broadly used for most cancers analysis and analysis. On this examine we examined the specificity of 4 commercially obtainable anti-p16INK4a antibodies in 4 immunological purposes.
The antibodies H-156 and JC8 detected the identical 16 kDa protein in western blot and immunoprecipitation assessments, whereas the antibody F-12 didn’t detect any protein in western blot evaluation or seize a protein that might be recognised by the H-156 antibody.
In immunocytochemistry assessments, the antibodies JC8 and H-156 detected a predominately cytoplasmic localised antigen, whose sign was depleted in p16INK4a siRNA experiments. F-12, in distinction, detected a predominately nuclear situated antigen and there was no noticeable discount on this sign after siRNA knockdown. Moreover in immunohistochemistry assessments, F-12 generated a special sample of staining in comparison with the JC8 and E6H4 antibodies.
These outcomes show that three out of 4 commercially obtainable p16INK4a antibodies are particular to, and point out a primarily cytoplasmic localisation for, the p16INK4a protein. The F-12 antibody, which has been broadly utilized in earlier research, gave totally different outcomes to the opposite antibodies and didn’t show specificity to human p16INK4a. This work emphasizes the significance of the validation of economic antibodies, apart to the beforehand reported use, for the complete verification of immunoreaction specificity.
Skinny HSIL of the Cervix: Detecting a Variant of Excessive-grade Squamous Intraepithelial Lesions With a p16INK4a Antibody.
The WHO defines skinny high-grade squamous intraepithelial lesions (HSIL) as a high-grade intraepithelial lesion of the cervix that’s often ≤9 cells thick. These lesions often develop in early metaplastic squamous epithelium with out anteceding low-grade squamous intraepithelial lesions (LSIL). The prevalence of skinny HSIL is just not effectively documented.
We evaluated totally different traits of skinny HSIL at time of remedy. We studied 25 formalin-fixed and paraffin-embedded conization specimens processed as step-serial sections. HSIL≤9 cells thick had been labeled as skinny HSIL. HSIL≥10 cells thick had been labeled as traditional HSIL. Immunohistochemical p16 staining was used to substantiate lesions of skinny HSIL.
General, 19 (76%) specimens contained each skinny HSIL and traditional HSIL, 4 (16%) contained skinny HSIL solely, 1 (4%) contained classic-type HSIL solely, and 1 (4%) contained skinny HSIL and LSIL. Skinny HSILs developed in each the columnar floor epithelium and deep cervical glandular epithelium.
Most skinny HSILs had been 5 cells thick. All HSILs (skinny and traditional) had been situated contained in the transformation zone and had a median horizontal extension of eight mm (vary, 0.Three to 21 mm). Our findings recommend that skinny HSILs are frequent findings, that they coexist with traditional HSIL, and ideally come up within the uncovered components of the transformation zone together with the glandular crypts.
An antibody to p16INK4A acknowledges a modified type of galectin-3.
Galectin-Three is a carbohydrate binding protein concerned in a number of processes together with cell-cycle regulation and apoptosis. The power of galectin-Three to guard cells from apoptosis depends upon a area of the protein often known as a BH-1 area for its homology to the anti-apoptotic protein Bcl-2. Right here, we present {that a} monoclonal antibody (MAb) to the human tumor suppressor protein p16INK4A acknowledges a post-translationally modified type of human galectin-3.
The modified type is detectable in solely a subset of cell varieties expressing galectin-3, indicating that the modification is cell-type-specific. Though there’s little amino acid sequence homology between p16INK4a and galectin-3, we present by epitope mapping that the modification straight impacts the construction of galectin-3’s BH-1 area. Elucidation of the character of this modification would possibly present additional perception into galectin-Three operate.
Using recombinant pseudotype virus-like particles harbouring inserted goal antigen to generate antibodies towards mobile marker p16INK4A.
Protein engineering gives a possibility to generate new immunogens with desired options. Beforehand, we’ve got demonstrated that hamster polyomavirus main capsid protein VP1-derived virus-like particles (VLPs) are extremely immunogenic and could be employed for the insertion of international epitopes at sure surface-exposed positions.
Within the present examine, we’ve got designed pseudotype VLPs consisting of an intact VP1 protein and VP2 protein fused with the goal antigen–cellular marker p16(INK4A)–at its N terminus. Each proteins coexpressed in yeast had been self-assembled to pseudotype VLPs harbouring the inserted antigen on the floor. The pseudotype VLPs had been used for era of antibodies towards p16(INK4A) that represents a possible biomarker for cells reworked by high-risk human papillomavirus (HPV).
The pseudotype VLPs induced in immunized mice a robust immune response towards the goal antigen. The antisera raised towards pseudotype VLPs confirmed particular immunostaining of p16(INK4A) protein in malignant cervical tissue. Spleen cells of the immunized mice had been used to generate monoclonal antibodies towards p16(INK4A) protein.
CDKN2A/p16INK4a Antibody |
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| AF0228 | Affbiotech | 200ul | EUR 420 |
p16INK4a Antibody / CDKN2A |
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| F54356-0.08ML | NSJ Bioreagents | 0.08 ml | EUR 140.25 |
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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, MDM1, 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. |
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p16INK4a Antibody / CDKN2A |
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| F54356-0.4ML | NSJ Bioreagents | 0.4 ml | EUR 322.15 |
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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, MDM1, 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. |
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p16INK4a Antibody / CDKN2A |
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| R20284-0.1ML | NSJ Bioreagents | 100ul | EUR 347.65 |
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Description: This recombinant p16INK4a antibody reacts to human p16INK4a (Cyclin-dependent kinase inhibitor 2A). |
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CDKN2A/p16INK4a Antibody |
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| AF5484-100ul | Affinity Biosciences | 100ul | EUR 280 |
CDKN2A/p16INK4a Antibody |
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| AF5484-200ul | Affinity Biosciences | 200ul | EUR 350 |
CDKN2A/p16INK4a Antibody |
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| AF6667-100ul | Affinity Biosciences | 100ul | EUR 280 |
CDKN2A/p16INK4a Antibody |
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| AF6667-200ul | Affinity Biosciences | 200ul | EUR 350 |
CDKN2A/p16INK4a Antibody |
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| AF6898-100ul | Affinity Biosciences | 100ul | EUR 280 |
CDKN2A/p16INK4a Antibody |
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| AF6898-200ul | Affinity Biosciences | 200ul | EUR 350 |
CDKN2A/p16INK4a Antibody |
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| AF5484 | Affinity Biosciences | 100ul | EUR 280 |
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Description: Human,Mouse |
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CDKN2A/p16INK4a Antibody |
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| AF6667 | Affinity Biosciences | 100ul | EUR 280 |
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Description: Human |
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CDKN2A/p16INK4a Antibody |
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| AF6898 | Affinity Biosciences | 100ul | EUR 280 |
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Description: Human |
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p16INK4a Antibody / CDKN2A |
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| V8604-100UG | NSJ Bioreagents | 100 ug | EUR 349.3 |
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Description: p16INK4a is a tumor suppressor protein. It is a specificinhibitor of cdk4/cdk6, and a tumorsuppressor involved inthe pathogenesis of a variety of malignancies. Recentanalyses of the p16INK4a gene revealed homozygousdeletions, nonsense, missense, or frameshift mutations inseveral human cancers. Although the frequency of p16INK4aabnormalities is higher in tumor derived cell lines than inunselected primary tumors, significant subsets of clinicalcases with aberrant p16INK4a gene have been reportedamong melanomas, gliomas, esophageal, pancreatic, lung,and urinary bladdercarcinomas, and some types of leukemia.Expression of p16INK4a (p16 positive) is highly correlatedwith human papilloma virus (HPV) infection in head and necksquamous cell carcinomas (HNSCC). p16 status is animportant prognostic indicator in HNSCC and the p16positive/HPV16 negative group is likely a distinct subgrouplacking any HPV genotype. |
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p16INK4a Antibody / CDKN2A |
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| V8604-20UG | NSJ Bioreagents | 20 ug | EUR 153.3 |
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Description: p16INK4a is a tumor suppressor protein. It is a specificinhibitor of cdk4/cdk6, and a tumorsuppressor involved inthe pathogenesis of a variety of malignancies. Recentanalyses of the p16INK4a gene revealed homozygousdeletions, nonsense, missense, or frameshift mutations inseveral human cancers. Although the frequency of p16INK4aabnormalities is higher in tumor derived cell lines than inunselected primary tumors, significant subsets of clinicalcases with aberrant p16INK4a gene have been reportedamong melanomas, gliomas, esophageal, pancreatic, lung,and urinary bladdercarcinomas, and some types of leukemia.Expression of p16INK4a (p16 positive) is highly correlatedwith human papilloma virus (HPV) infection in head and necksquamous cell carcinomas (HNSCC). p16 status is animportant prognostic indicator in HNSCC and the p16positive/HPV16 negative group is likely a distinct subgrouplacking any HPV genotype. |
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p16INK4a Antibody / CDKN2A |
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| V8604SAF-100UG | NSJ Bioreagents | 100 ug | EUR 349.3 |
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Description: p16INK4a is a tumor suppressor protein. It is a specificinhibitor of cdk4/cdk6, and a tumorsuppressor involved inthe pathogenesis of a variety of malignancies. Recentanalyses of the p16INK4a gene revealed homozygousdeletions, nonsense, missense, or frameshift mutations inseveral human cancers. Although the frequency of p16INK4aabnormalities is higher in tumor derived cell lines than inunselected primary tumors, significant subsets of clinicalcases with aberrant p16INK4a gene have been reportedamong melanomas, gliomas, esophageal, pancreatic, lung,and urinary bladdercarcinomas, and some types of leukemia.Expression of p16INK4a (p16 positive) is highly correlatedwith human papilloma virus (HPV) infection in head and necksquamous cell carcinomas (HNSCC). p16 status is animportant prognostic indicator in HNSCC and the p16positive/HPV16 negative group is likely a distinct subgrouplacking any HPV genotype. |
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p16INK4a Antibody / CDKN2A |
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| V8694-100UG | NSJ Bioreagents | 100 ug | EUR 349.3 |
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Description: p16INK4a is a tumor suppressor protein. It is a specific inhibitor of cdk4/cdk6, and a tumor suppressor involved in the pathogenesis of a variety of malignancies. Recent analyses of the p16INK4a gene revealed homozygous deletions, nonsense, missense, or frameshift mutations in several human cancers. Although the frequency of p16INK4a abnormalities is higher in tumor derived cell lines than in unselected primary tumors, significant subsets of clinical cases with aberrant p16INK4a gene have been reported among melanomas, gliomas, esophageal, pancreatic, lung, and urinary bladder carcinomas, and some types of leukemia. Expression of p16INK4a (p16 positive) is highly correlated with human papilloma virus (HPV) infection in head and neck squamous cell carcinomas (HNSCC). p16 status is an important prognostic indicator in HNSCC and the p16 positive/HPV16 negative group is likely a distinct subgroup lacking any HPV genotype. |
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p16INK4a Antibody / CDKN2A |
|||
| V8694-20UG | NSJ Bioreagents | 20 ug | EUR 153.3 |
|
Description: p16INK4a is a tumor suppressor protein. It is a specific inhibitor of cdk4/cdk6, and a tumor suppressor involved in the pathogenesis of a variety of malignancies. Recent analyses of the p16INK4a gene revealed homozygous deletions, nonsense, missense, or frameshift mutations in several human cancers. Although the frequency of p16INK4a abnormalities is higher in tumor derived cell lines than in unselected primary tumors, significant subsets of clinical cases with aberrant p16INK4a gene have been reported among melanomas, gliomas, esophageal, pancreatic, lung, and urinary bladder carcinomas, and some types of leukemia. Expression of p16INK4a (p16 positive) is highly correlated with human papilloma virus (HPV) infection in head and neck squamous cell carcinomas (HNSCC). p16 status is an important prognostic indicator in HNSCC and the p16 positive/HPV16 negative group is likely a distinct subgroup lacking any HPV genotype. |
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p16INK4a Antibody / CDKN2A |
|||
| V8694SAF-100UG | NSJ Bioreagents | 100 ug | EUR 349.3 |
|
Description: p16INK4a is a tumor suppressor protein. It is a specific inhibitor of cdk4/cdk6, and a tumor suppressor involved in the pathogenesis of a variety of malignancies. Recent analyses of the p16INK4a gene revealed homozygous deletions, nonsense, missense, or frameshift mutations in several human cancers. Although the frequency of p16INK4a abnormalities is higher in tumor derived cell lines than in unselected primary tumors, significant subsets of clinical cases with aberrant p16INK4a gene have been reported among melanomas, gliomas, esophageal, pancreatic, lung, and urinary bladder carcinomas, and some types of leukemia. Expression of p16INK4a (p16 positive) is highly correlated with human papilloma virus (HPV) infection in head and neck squamous cell carcinomas (HNSCC). p16 status is an important prognostic indicator in HNSCC and the p16 positive/HPV16 negative group is likely a distinct subgroup lacking any HPV genotype. |
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P16INK4A Antibody (IHC Gold) |
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| E36PA116 | EnoGene | 100ul | EUR 225 |
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Description: Available in various conjugation types. |
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Anti-CDKN2A/p16INK4a Antibody |
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| ER1803-53 | HUABIO | 100ul | EUR 189 |
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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. |
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Anti-CDKN2A/p16INK4a Antibody |
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| ER2001-30 | HUABIO | 100ul | EUR 189 |
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Description: CDKN2A, also known as cyclin-dependent kinase inhibitor 2A, is a gene which in humans is located at chromosome 9, band p21.3. It is ubiquitously expressed in many tissues and cell types.The gene codes for two proteins, including the INK4 family member p16 (or p16INK4a) and p14arf. Both act as tumor suppressors by regulating the cell cycle. p16 inhibits cyclin dependent kinases 4 and 6 (CDK4 and CDK6) and thereby activates the retinoblastoma (Rb) family of proteins, which block traversal from G1 to S-phase. p14ARF (known as p19ARF in the mouse) activates the p53 tumor suppressor. Somatic mutations of CDKN2A are common in the majority of human cancers, with estimates that CDKN2A is the second most commonly inactivated gene in cancer after p53. Germline mutations of CDKN2A are associated with familial melanoma, glioblastoma and pancreatic cancer.The CDKN2A gene also contains one of 27 SNPs associated with increased risk of coronary artery disease. |
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The specificity of antibodies was confirmed by the immunostaining of HPV-transformed cells. In conclusion, the present examine demonstrates the potential of pseudotype VLPs with inserted goal antigen as a brand new kind of immunogens to generate antibodies of excessive diagnostic worth.
