Results: Yeager et al. Thermo Fisher Scientific. Tests used on biopsy and cytology specimens to diagnose cancer Xing SS,Li J,Chen L,Yang YF,He PL,Li J,Yang J. Zhang JJ,Zhang YZ,Peng JJ,Li NS,Xiong XM,Ma QL,Luo XJ,Liu B,Peng J. The p16 protein has been identified as a specific inhibitor of cdk4 because it blocks cdk4 substrate phosphorylation. Do not freeze. Molecular pathways: cDK4 inhibitors for cancer therapy. Wang S,Cao Z,Xue J,Li H,Jiang W,Cheng Y,Li G,Zhang X. Potency of Human Cardiosphere-Derived Cells from Patients with Ischemic Heart Disease Is Associated with Robust Vascular Supportive Ability. J Clin Pathol. A new regulatory motif in cell-cycle control causing specific inhibition of cyclin D/CDK4. Discard supernatant in appropriate waste container. 550337) as the secondary antibody and Streptavidin-HRP (Cat. Approximately 10,000 cells can be analyzed and processed by a computer in less than one minute. Gynecol Oncol. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated p16 INK4A (D7C1M) Rabbit mAb #80772. Iron accumulation in senescent cells is coupled with impaired ferritinophagy and inhibition of ferroptosis. Flow Cytometry, Methanol Permeabilization Protocol for Directly Conjugated Antibodies A. Chemopreventive efficacy of salvianolic acid B phospholipid complex loaded nanoparticles against experimental oral carcinogenesis: implication of sustained drug release. p16 INK4A (D7C1M) Rabbit mAb (Alexa Fluor - Cell Signaling Technology Wash cells by centrifugation in excess 1X PBS to remove methanol. documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any Members of the INK4 family of cyclin dependent kinase inhibitors include p16 INK4A, p15 INK4B, p18 INK4C, and p19 INK4D. Titration of the reagents or substituting with other (non-matched) isotype control is NOT recommended. Cells expressing p16 were visualized using a three-step staining procedure with Polyclonal Biotin Goat Anti-Mouse Ig (Cat. commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying 550878). Not for resale without express authorization. No. NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to fixation. In contrast, hybridoma-based systems for producing monoclonal antibodies are subject to genetic Not for use in diagnostic procedures. ZNRD1-AS1 knockdown alleviates malignant phenotype of retinoblastoma through miR-128-3p/BMI1 axis. Cells in e. analyzed for mitochondrial superoxide by flow cytometry. scientific content. The p16INK4A -abnormal situation was an independent HSIL+ risk factor for 2-year outcomes in all three cervicopathological conditions investigated (hazard ratios: 4.3-7.2). Resuspend cells in 100 l of diluted primary antibody, prepared in Antibody Dilution Buffer at a recommended dilution or as determined via titration. STRING - Known and Predicted Protein-Protein Interactions. No. A comparative analysis using abnormal cervical smears with follow-up biopsies. Would you like to stay on the current country site or be switched to your country? Recombinant antibodies offer several key advantages compared to traditional antibodies. Science. All reagents required for this protocol may be efficiently purchased together in our Intracellular Flow cervical cancer; flow cytometry; human papillomavirus; immunostaining; p16INK4A; papanicolaou smear; pathology; quantification; squamous intraepithelial lesion. All rights reserved. Store at 4C. p16 Antibody, anti-human, REAfinity | Recombinant antibodies | MACS NOTE: Antibodies targeting CD markers or other extracellular proteins may be added prior to fixation if the epitope is disrupted by formaldehyde and/or methanol. Objective: To investigate DNA aneuploid and P16 expression in biopsy specimens from lung cancer, and to study genetic instability and the application of flow cytometry in lung cancer pernicious degree diagnosis. -, Senapati R, Senapati NN, Dwibedi B. Molecular mechanisms of HPV mediated neoplastic progression. (A) Examination of, The research profile of twotier clinical validation work for p16 INK4A FCM. (Biology). Not for resale without express authorization. BD Pharmingen Purified Mouse Anti-Human p16. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use. waste container. Flow cytometric analysis was performed on a BD FACScan. Permeabilize for a minimum of 10 min on ice. secreted by many different B cell clones and recognize multiple antigenic epitopes, monoclonals originate from a single B Biochemistry (Mosc). 2022 Apr 27;22(1):138. doi: 10.1186/s12905-022-01714-0. Wash by centrifugation with excess 1X PBS. Lazzarini E,Balbi C,Altieri P,Pfeffer U,Gambini E,Canepa M,Varesio L,Bosco MC,Coviello D,Pompilio G,Brunelli C,Cancedda R,Ameri P,Bollini S. HBP1-mediated Regulation of p21 Protein through the Mdm2/p53 and TCF4/EZH2 Pathways and Its Impact on Cell Senescence and Tumorigenesis. 2005 Apr;97(1):35-40. doi: 10.1016/j.ygyno.2004.11.034. Analyte Specific Reagent. Link to hereditary melanoma brightens mood for p16 gene. or blood specimen is treated with special antibodies. ARF; CDK4I; CDKN2; CDKN2A; CMM2; INK4; INK4A; MLM; MTS-1; MTS1; P14; P14ARF; P16; P16-INK4A; P16INK4; P16INK4A; P19; P19ARF; TP16, If an Invitrogen antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*. Clicking the images or links will redirect you to a website hosted by BenchSci that provides third-party Journal of assisted reproduction and genetics, Yan P,Xu J,Zeng Y,Dong G,Cao H,Zheng M,Zhu H. Knockout of 4.1B triggers malignant transformation in SV40T-immortalized mouse embryo fibroblast cells. apply to Products provided by CST, its affiliates or its distributors. Discard supernatant in appropriate waste container. In cross-sectional studies, age- and viral genotype-dependent expression of p16INK4A was investigated, and optimal diagnostic parameter cut-offs (using colposcopy and biopsy as a gold standard) were obtained. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. FCM-based p16INK4A quantification offers a better choice for conveniently and precisely monitoring the occurrence of HSIL+ and directing risk-stratification-based interventions. Using this assay, we were able to quantify the activity level of SA -galactosidase, the expression level of p16 INK4a and H2AX in these cell populations. Recently a gene cloned from the short arm of human chromosome 9, Multiple Tumor Suppressor 1 (MTS1) has been identified as the gene for p16. Motterle A,Pu X,Wood H,Xiao Q,Gor S,Ng FL,Chan K,Cross F,Shohreh B,Poston RN,Tucker AT,Caulfield MJ,Ye S, Copyright 2006-2023 Thermo Fisher Scientific Inc. All rights reserved. CRB3 regulates contact inhibition by activating the Hippo pathway in mammary epithelial cells. Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable cells. Epub 2014 May 12. Mix well to dissociate pellet and prevent cross-linking of individual cells. To use this carrier-free antibody for conjugation experiments, we strongly recommend performing another round of desalting. Aromseree S, Wongjumpa W, Ekalaksananan T, Temtanakitpaisan A, Kleebkaow P, Srisathaporn S, Tongchai P, Pientong C. Asian Pac J Cancer Prev. the tissue culture supernatants of transfected host cell lines. Stojiljkovic MR,Ain Q,Bondeva T,Heller R,Schmeer C,Witte OW. addressing the ongoing reproducibility crisis. are provided for Customer as the end-user and solely for research and development uses. The INK4 family members inhibit cyclin dependent kinases 4 and 6 (CDK4 and CDK6), causing cell cycle arrest in G1 phase. Alternatively, cells may be stored overnight at 4C in 1X PBS. The site you are about to visit is operated by a third party. p16 INK4A (D7C1M) Rabbit mAb (Alexa Fluor 647 Conjugate), This Cell Signaling Technology antibody is conjugated to Alexa Fluor. The gene, now also known as the CDKN2 gene, has been found to be mutated in a very high percentage of tumors, including 75% of melanoma cell lines. Bethesda, MD 20894, Web Policies Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed Murine mesenchymal cells that express elevated levels of the CDK inhibitor p16(Ink4a) in vivo are not necessarily senescent. p16 INK4A protein expression is often altered in human cancer, and high expression is currently used as a predictive biomarker in cervical cancer (3-5). Xu X,Pang J,Chen Y,Bucala R,Zhang Y,Ren J. Source of all serum proteins is from USDA inspected abattoirs located in the United States. For analysis by flow cytometry, peritoneal cells were washed with HBSS plus 2% heat-inactivated FBS and blocked with anti-CD16/CD32 (clone 93) for 10 min on ice . 2015 Jun 15;136(12):2741-51. doi: 10.1002/ijc.28900. Alternate names for p16 include. Teng-Long-Bu-Zhong-Tang, a Chinese herbal formula, enhances anticancer effects of 5--Fluorouracil in CT26 colon carcinoma. We offer antibodies conjugated to many types of fluorophores to accommodate your flow cytometry needs. Bergeron C, Ronco G, Reuschenbach M, Wentzensen N, Arbyn M, Stoler M, von Knebel Doeberitz M. Int J Cancer. selection have been evaluated by us; we are providing them as-is and without warranty of any kind, Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry. All rights reserved. View specifications, prices, citations, reviews, and more. We have developed an alternative flow cytometry-based method that simultaneously quantifies multiple senescence markers at a single-cell resolution. Comparative characterization of SHED and DPSCs during extended cultivation invitro. (Immunogen). 550878). Identification of High-Risk Human Papillomavirus DNA, p16, and E6/E7 Oncoproteins in Laryngeal and Hypopharyngeal Squamous Cell Carcinomas. requires a separate license from CST. Proceed with immunostaining (Section D) or store cells at -20C in 90% methanol. All reagents required for this protocol may be efficiently purchased together in our Intracellular Flow Herein, we created a high-throughput, quantitative diagnostic device, p16INK4A flow cytometry (FCM) and assessed its performances in cervical cancer screening and prevention. The p16 protein has been identified as a specific inhibitor of cdk4 because it blocks cdk4 substrate phosphorylation. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala143 of human p16 INK4A protein. lines of bacterial, yeast, or insect origin are also suitable. Heart and circulatory physiology, Li Y,Peng Z,Wang C,Li L,Leng Y,Chen R,Yuan H,Zhou S,Zhang Z,Chen AF. Not for use in diagnostic procedures. Protect from light. The gene, now also known as the CDKN2 gene, has been found to be mutated in a very high percentage of tumors, including 75% of melanoma cell lines. Anti-P16 Antibody Products | Biocompare PMC 2022 The Authors. This antibody does not cross-react with p15 INK4B. Build your flow cytometry panels with our antibodies to identify cells and detect proliferation, homing profiles, activation states, and cytokine release. addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized After cloning the antibody genes into an expression vector, this is then transfected into an appropriate host cell line Product Details Product Specific Information Clin Cancer Res. Immunohistochemistry:The p16 antibody is recommended to test for immunohistochemical staining on formalin-fixed paraffin-embedded sections. For optimal indirect immunohistochemical staining, this p16 antibody should be titrated (1:10 to 1:50 dilution) and visualized via a three-step staining procedure in combination with the Polyclonal Biotin Goat Anti-Mouse Ig (Cat. and 1:500 Anti-CDKN2A/p16 INK4a (EPR1473, Abcam, ab209579) antibody was added and incubated at room temperature for 30 min in the dark. Nieh S, Chen SF, Chu TY, Lai HC, Lin YS, Fu E, Gau CH. Permeabilized WI-26 VA4 cells were stained with either FITC Mouse Anti-Human p16 (Component No. Deficiency of insulin-like growth factor 1 attenuates aging-induced changes in hepatic function: role of autophagy. The authors declare no conflicts of interest. 10883-1-AP reacts with p16 specifically. Chen X,Liu C,Zhao R,Zhao P,Wu J,Zhou N,Ying M. Analysis of the relationship between the KRAS G12V oncogene and the Hippo effector YAP1 in embryonal rhabdomyosarcoma. cell clone and are specific for just one epitope. Permeabilized WI-26 VA4 cells were stained with either FITC Mouse Anti-Human p16 (Component No. Reciprocal Rb inactivation and p16INK4 expression in primary lung cancers and cell lines.