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 244

p16INK4A Conjugated Antibody

C48843 100ul
EUR 397

Anti-P16INK4a antibody

STJ180203 0.1 ml
EUR 212

Anti-P16INK4a antibody

STJ180271 0.1 ml
EUR 215

Anti-P16INK4a antibody

STJ180410 0.1 ml
EUR 212

CDKN2A / p16INK4a Polyclonal Antibody

27512-100ul 100ul
EUR 252

CDKN2A / p16INK4a Polyclonal Antibody

27512-50ul 50ul
EUR 187

CDKN2A / p16INK4a Polyclonal Antibody

27527-100ul 100ul
EUR 252

CDKN2A / p16INK4a Polyclonal Antibody

27527-50ul 50ul
EUR 187

Polyclonal CDKN2A / p16INK4a Antibody

APR02292G 0.05ml
EUR 484
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human CDKN2A / p16INK4a . This antibody is tested and proven to work in the following applications:

Anti-CDKN2A / p16INK4a antibody

STJ11100176 100 µl
EUR 457
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.

Antibody for Human p16INK4a

SPC-1280D 0.1ml
EUR 314
  • p16INK4a is a 16 kDa member of the CDKN2 cyclin-dependent kinase inhibitor family of molecules. It is widely expressed (although not in skeletal muscle) and serves as a negative regulator of cell proliferation. It does so by associating with CDK4 or
  • Show more
Description: A polyclonal antibody for p16INK4a from Human | Mouse. The antibody is produced in rabbit after immunization with Human Synthesized peptide derived from human p12 INK antibody.. The Antibody is tested and validated for WB, ELISA assays with the following recommended dilutions: WB (1:1000), ELISA (1:5000). This p16INK4a antibody is unconjugated.

Antibody for Human p16INK4a

SPC-1280D-A390 0.1ml
EUR 361
  • p16INK4a is a 16 kDa member of the CDKN2 cyclin-dependent kinase inhibitor family of molecules. It is widely expressed (although not in skeletal muscle) and serves as a negative regulator of cell proliferation. It does so by associating with CDK4 or
  • Show more
Description: A polyclonal antibody for p16INK4a from Human | Mouse. The antibody is produced in rabbit after immunization with Human Synthesized peptide derived from human p12 INK antibody.. The Antibody is tested and validated for WB, ELISA assays with the following recommended dilutions: WB (1:1000), ELISA (1:5000). This p16INK4a antibody is conjugated to ATTO 390.

Antibody for Human p16INK4a

SPC-1280D-A488 0.1ml
EUR 360
  • p16INK4a is a 16 kDa member of the CDKN2 cyclin-dependent kinase inhibitor family of molecules. It is widely expressed (although not in skeletal muscle) and serves as a negative regulator of cell proliferation. It does so by associating with CDK4 or
  • Show more
Description: A polyclonal antibody for p16INK4a from Human | Mouse. The antibody is produced in rabbit after immunization with Human Synthesized peptide derived from human p12 INK antibody.. The Antibody is tested and validated for WB, ELISA assays with the following recommended dilutions: WB (1:1000), ELISA (1:5000). This p16INK4a antibody is conjugated to ATTO 488.

Antibody for Human p16INK4a

SPC-1280D-A565 0.1ml
EUR 360
  • p16INK4a is a 16 kDa member of the CDKN2 cyclin-dependent kinase inhibitor family of molecules. It is widely expressed (although not in skeletal muscle) and serves as a negative regulator of cell proliferation. It does so by associating with CDK4 or
  • Show more
Description: A polyclonal antibody for p16INK4a from Human | Mouse. The antibody is produced in rabbit after immunization with Human Synthesized peptide derived from human p12 INK antibody.. The Antibody is tested and validated for WB, ELISA assays with the following recommended dilutions: WB (1:1000), ELISA (1:5000). This p16INK4a antibody is conjugated to ATTO 565.

Antibody for Human p16INK4a

SPC-1280D-A594 0.1ml
EUR 360
  • p16INK4a is a 16 kDa member of the CDKN2 cyclin-dependent kinase inhibitor family of molecules. It is widely expressed (although not in skeletal muscle) and serves as a negative regulator of cell proliferation. It does so by associating with CDK4 or
  • Show more
Description: A polyclonal antibody for p16INK4a from Human | Mouse. The antibody is produced in rabbit after immunization with Human Synthesized peptide derived from human p12 INK antibody.. The Antibody is tested and validated for WB, ELISA assays with the following recommended dilutions: WB (1:1000), ELISA (1:5000). This p16INK4a antibody is conjugated to ATTO 594.

Antibody for Human p16INK4a

SPC-1280D-A633 0.1ml
EUR 360
  • p16INK4a is a 16 kDa member of the CDKN2 cyclin-dependent kinase inhibitor family of molecules. It is widely expressed (although not in skeletal muscle) and serves as a negative regulator of cell proliferation. It does so by associating with CDK4 or
  • Show more
Description: A polyclonal antibody for p16INK4a from Human | Mouse. The antibody is produced in rabbit after immunization with Human Synthesized peptide derived from human p12 INK antibody.. The Antibody is tested and validated for WB, ELISA assays with the following recommended dilutions: WB (1:1000), ELISA (1:5000). This p16INK4a antibody is conjugated to ATTO 633.

Antibody for Human p16INK4a

SPC-1280D-A655 0.1ml
EUR 360
  • p16INK4a is a 16 kDa member of the CDKN2 cyclin-dependent kinase inhibitor family of molecules. It is widely expressed (although not in skeletal muscle) and serves as a negative regulator of cell proliferation. It does so by associating with CDK4 or
  • Show more
Description: A polyclonal antibody for p16INK4a from Human | Mouse. The antibody is produced in rabbit after immunization with Human Synthesized peptide derived from human p12 INK antibody.. The Antibody is tested and validated for WB, ELISA assays with the following recommended dilutions: WB (1:1000), ELISA (1:5000). This p16INK4a antibody is conjugated to ATTO 655.

Antibody for Human p16INK4a

SPC-1280D-A680 0.1ml
EUR 360
  • p16INK4a is a 16 kDa member of the CDKN2 cyclin-dependent kinase inhibitor family of molecules. It is widely expressed (although not in skeletal muscle) and serves as a negative regulator of cell proliferation. It does so by associating with CDK4 or
  • Show more
Description: A polyclonal antibody for p16INK4a from Human | Mouse. The antibody is produced in rabbit after immunization with Human Synthesized peptide derived from human p12 INK antibody.. The Antibody is tested and validated for WB, ELISA assays with the following recommended dilutions: WB (1:1000), ELISA (1:5000). This p16INK4a antibody is conjugated to ATTO 680.

Antibody for Human p16INK4a

SPC-1280D-A700 0.1ml
EUR 360
  • p16INK4a is a 16 kDa member of the CDKN2 cyclin-dependent kinase inhibitor family of molecules. It is widely expressed (although not in skeletal muscle) and serves as a negative regulator of cell proliferation. It does so by associating with CDK4 or
  • Show more
Description: A polyclonal antibody for p16INK4a from Human | Mouse. The antibody is produced in rabbit after immunization with Human Synthesized peptide derived from human p12 INK antibody.. The Antibody is tested and validated for WB, ELISA assays with the following recommended dilutions: WB (1:1000), ELISA (1:5000). This p16INK4a antibody is conjugated to ATTO 700.

Antibody for Human p16INK4a

SPC-1280D-ALP 0.1ml
EUR 355
  • p16INK4a is a 16 kDa member of the CDKN2 cyclin-dependent kinase inhibitor family of molecules. It is widely expressed (although not in skeletal muscle) and serves as a negative regulator of cell proliferation. It does so by associating with CDK4 or
  • Show more
Description: A polyclonal antibody for p16INK4a from Human | Mouse. The antibody is produced in rabbit after immunization with Human Synthesized peptide derived from human p12 INK antibody.. The Antibody is tested and validated for WB, ELISA assays with the following recommended dilutions: WB (1:1000), ELISA (1:5000). This p16INK4a antibody is conjugated to Alkaline Phosphatase.

Antibody for Human p16INK4a

SPC-1280D-APC 0.1ml
EUR 359
  • p16INK4a is a 16 kDa member of the CDKN2 cyclin-dependent kinase inhibitor family of molecules. It is widely expressed (although not in skeletal muscle) and serves as a negative regulator of cell proliferation. It does so by associating with CDK4 or
  • Show more
Description: A polyclonal antibody for p16INK4a from Human | Mouse. The antibody is produced in rabbit after immunization with Human Synthesized peptide derived from human p12 INK antibody.. The Antibody is tested and validated for WB, ELISA assays with the following recommended dilutions: WB (1:1000), ELISA (1:5000). This p16INK4a antibody is conjugated to APC .
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

Monoclonal antibodies to fetal bovine serum acetylcholinesterase distinguish between acetylcholinesterases from ruminant and non-ruminant species

Monoclonal antibodies to fetal bovine serum acetylcholinesterase distinguish between acetylcholinesterases from ruminant and non-ruminant speciesMonoclonal antibodies to fetal bovine serum acetylcholinesterase distinguish between acetylcholinesterases from ruminant and non-ruminant species

Two kinds of cholinesterases (ChEs) are current in mammalian blood and tissues: acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Whereas AChE regulates neurotransmission by hydrolyzing acetylcholine on the postsynaptic membranes and neuromuscular junctions, BChE in plasma