Immunohistochemical p16INK4a analysis of archival tumors with deletion, hypermethylation, or mutation of the CDKN2/MTS1 gene. A comparison of four commercial antibodies.

Immunohistochemical p16INK4a analysis of archival tumors with deletion, hypermethylation, or mutation of the CDKN2/MTS1 gene. A comparison of four commercial antibodies.
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.

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.

P16INK4A

MO47002 100 ul
EUR 292.8

p16INK4A Conjugated Antibody

C48843 100ul
EUR 476.4

CDKN2A/p16INK4a Antibody

AF5484 200ul
EUR 420

CDKN2A/p16INK4a Antibody

AF6667 100ul
EUR 420

CDKN2A/p16INK4a Antibody

AF6898 100ul
EUR 420

CDKN2A/p16INK4a Antibody

AF0228 200ul
EUR 420

Anti-P16INK4a antibody

STJ180203 0.1 ml
EUR 254.4

Anti-P16INK4a antibody

STJ180271 0.1 ml
EUR 258

Anti-P16INK4a antibody

STJ180410 0.1 ml
EUR 254.4

p16INK4a Antibody / CDKN2A

F54356-0.08ML 0.08 ml
EUR 165

p16INK4a Antibody / CDKN2A

F54356-0.4ML 0.4 ml
EUR 379

p16INK4a Antibody / CDKN2A

R20284-0.1ML 100ul
EUR 409

p16INK4a Antibody / CDKN2A

V8604-100UG 100 ug
EUR 499
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.

p16INK4a Antibody / CDKN2A

V8604-20UG 20 ug
EUR 219
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.

p16INK4a Antibody / CDKN2A

V8604SAF-100UG 100 ug
EUR 499
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.

p16INK4a Antibody / CDKN2A

V8694-100UG 100 ug
EUR 499
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.

p16INK4a Antibody / CDKN2A

V8694-20UG 20 ug
EUR 219
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.

p16INK4a Antibody / CDKN2A

V8694SAF-100UG 100 ug
EUR 499
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.

CDKN2A / p16INK4a Polyclonal Antibody

27512-100ul 100ul
EUR 302.4

CDKN2A / p16INK4a Polyclonal Antibody

27512-50ul 50ul
EUR 224.4

CDKN2A / p16INK4a Polyclonal Antibody

27527-100ul 100ul
EUR 302.4

CDKN2A / p16INK4a Polyclonal Antibody

27527-50ul 50ul
EUR 224.4
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.

Leave a Reply

Your email address will not be published. Required fields are marked *

Related Post

Isolation and characterization of a novel GRP78-specific single-chain variable fragment (scFv) using ribosome display method

Isolation and characterization of a novel GRP78-specific single-chain variable fragment (scFv) using ribosome display methodIsolation and characterization of a novel GRP78-specific single-chain variable fragment (scFv) using ribosome display method

Glucose-regulated protein 78 (GRP78) is a well-characterized endoplasmic reticulum (ER) chaperon steadily overexpressed on the floor of tumor cells and related to tumor survival, metastasis, and chemoresistance. Therefore, potential GRP78