Anti p16

Manufactured by Abcam

Sourced in United States, United Kingdom

Anti-p16 is a primary antibody that specifically binds to the p16 protein. p16 is a tumor suppressor protein that inhibits cell cycle progression. This antibody can be used in various laboratory techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to detect and analyze the expression of p16 in biological samples.

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Lab products found in correlation

Anti p21, by Abcam (8 mentions) Anti p53, by Abcam (5 mentions) Anti β actin, by Santa Cruz Biotechnology (3 mentions) Bca protein assay kit, by Thermo Fisher Scientific (3 mentions) Pvdf membrane, by Merck Group (3 mentions) Anti sirt1, by Abcam (3 mentions) Anti p21, by Santa Cruz Biotechnology (3 mentions) Anti p53, by Santa Cruz Biotechnology (2 mentions) Anti flag, by Merck Group (2 mentions) Anti p akt1, by Abcam (2 mentions) Anti bcl 2, by Abcam (2 mentions) Anti gapdh, by Abcam (2 mentions) Anti cyclin d1, by Cell Signaling Technology (2 mentions) Anti p21, by Cell Signaling Technology (2 mentions) Ripa lysis buffer, by Beyotime (2 mentions) Anti gapdh, by Cell Signaling Technology (2 mentions) Anti p21, by Proteintech (2 mentions) Anti gapdh, by Proteintech (2 mentions) Polyvinylidene fluoride membrane, by Merck Group (2 mentions) Anti rb, by Abcam (2 mentions)

Close spelling variants of this product

Anti-P16 antibody (4 citations) Anti-p16 ARC (mAb EP1551Y) (3 citations) Rabbit anti-p16 (3 citations) Rabbit anti-human p16 (2 citations) Anti‐ ‐mouse p16 (2 citations) Rabbit anti-p16 antibody (2 citations)

32 protocols using anti p16

Molecular Mechanisms of Senescence in Neurons

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PFT-α was obtained from Sigma-Aldrich China (Shanghai, China; P4236). The TBK inhibitor BX-795 and the Akt inhibitor MK-2206 were purchased from Selleck chemicals (Houston, TX, USA) (s1274 and s1078). Senescence β-gal staining kit (Cell Signaling Technology, Beverly, MA, USA; #9860).
The following antibodies were used in western blot: anti-TBK1 (Abcam, Cambridge, MA, USA, ab-40676, 1:1000), anti-p16 (Abcam, ab-51243, 1:1000), and anti-cyclin A (Santa Cruz, Dallas, TX, USA; sc-596, 1:1000), anti-pAkt 473 (Cell Signaling Technology; #4060, 1:2000), anti-Bmi (Cell Signaling Technology; #648, 1:1000), anti-phosphorylation serine (Boster, China, BM1622), anti-p53 (Santa Cruz; sc-126, 1:1000), anti-β-actin (Santa Cruz; sc-47778, 1:1000). The following antibodies were used in immunofluorescence: anti-NeuN (Abcam, ab-177487, 1:200), anti-Brn-3a (Santa Cruz; sc-8429, 1:200), anti-p16 (Abcam, ab-51243, 1:100), anti-TBK1 (Abcam, ab-40676, 1:200). anti-pAkt 473 (Cell Signaling Technology; #4060, 1:200) was used in immunohistochemistry.

Li L.U., Zhao Y, & Zhang H. (2017). P16INK4a upregulation mediated by TBK1 induces retinal ganglion cell senescence in ischemic injury. Cell Death & Disease, 8(4), e2752-.

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Protein Lysate Extraction and Western Blotting

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Whole-cell protein lysates were extracted from cells by lysis with NP250 buffer (20 mM Tris (pH 7.6), 1 mM EDTA, 0.5% NP40 and 235 mM NaCl). In gene silencing experiments, cells were cultured for 48 h after siRNA transfection, at which point lysates were generated. Following a 20 min incubation at 4 °C, lysates were centrifuged and supernatents collected. Electrophoresis was performed using Novex precast Bis-Tris gels (Invitrogen) and gels were blotted onto nitrocellulose filters as described previously [64 (link)]. Blots were immunoblotted in 5% (w/v) milk at 4 °C overnight using the following primary antibodies: anti-Rb1 (1/1000 (v/v) dilution in 5% (v/v) milk, New England Biolab, 9309); anti-p16 (1/1000, abcam); anti-SKP2 (1/1000, New England Biolab, 4358); anti-p27 (1/1000, New Engand Biolab, 2552); anti-tubulin (1/1000, abcam); and anti-actin (1/1000, Santa Cruz, sc-1616). After washing, blots were incubated 1 h at room temperature with secondary antibodies (Li-COR) diluted 1/10 000 (v/v) in 5% (w/v) milk. Protein bands were visualised and quantified using the Odyssey FC imaging system and ImageStudio software (Li-COR).

Brough R., Gulati A., Haider S., Kumar R., Campbell J., Knudsen E., Pettitt S.J., Ryan C.J, & Lord C.J. (2018). Identification of highly penetrant Rb-related synthetic lethal interactions in triple negative breast cancer. Oncogene, 37(43), 5701-5718.

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Immunoblotting for Protein Analysis

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Immunoblotting was performed as previously described [22 (link)]. Anti-Flag (Cat. F7425) was purchased from Sigma. Anti-GAPDH (Cat. sc-47724) was purchased from Santa Cruz Biotech. Anti-Jarid2 (Cat. ab48137) and anti-p16 (Cat. ab201980) were purchased from Abcam.

Zhu X.X., Yan Y.W., Ai C.Z., Jiang S., Xu S.S., Niu M., Wang X.Z., Zhong G.S., Lu X.F., Xue Y., Tian S., Li G., Tang S, & Jiang Y.Z. (2017). Jarid2 is essential for the maintenance of tumor initiating cells in bladder cancer. Oncotarget, 8(15), 24483-24490.

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Western Blot Analysis of Cancer Markers

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Cell lysates were collected in 1X sample buffer (2% SDS, 10% glycerol, 0.01% bromophenol blue, 62.5mM Tris, pH=6.8, 0.1M DTT) and boiled (10 min, 95° C). Protein concentration was determined using Bradford assay (Bio-Rad, cat#5000006). Proteins were resolved using SDS-PAGE gels and transferred to nitrocellulose membranes (GE Healthcare Life Sciences, cat#10600001) as previously described [8 (link)]. Antibodies used include: anti-BRAF (Santa Cruz Biotechnology, cat#sc-5284, 1:1000), anti-RAS (BD Sciences, cat#610001, 1:1000), anti-p16 (Abcam, cat#ab108349, 1:1000), anti-p21 (Abcam cat#ab109199, 1:1000), anti-cyclin A2 (Abcam cat#ab181591, 1:2000), anti-vinculin (Sigma-Aldrich cat#V9131, 1:1000), β-Actin (Sigma-Aldrich, cat#A1978, 1:10000), anti-mouse HRP (Cell Signaling Technology, cat#cst7076, 1:10,000), and anti-rabbit HRP (Cell Signaling Technology, cat#cst7074, 1:5000).

Buj R., Leon K.E., Anguelov M.A, & Aird K.M. (2021). Suppression of p16 alleviates the senescence-associated secretory phenotype. Aging (Albany NY), 13(3), 3290-3312.

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Antibody-based Expression Analysis in Cancer Cells

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The following antibodies were used for immunofluorescence staining: anti-GAPDH (1:5,000; cat. no. Ab8245; Abcam); anti-UCP2 (1:100; cat. no. 11081-1-AP; ProteinTech Group, Inc.) and anti-FLNa (1:100; cat. no. ab76289; Abcam). The antibodies were used to detect UCP2 and FLNa expression in different CC cell lines.
The following antibodies were used for immunohistochemical staining: anti-UCP2 (1:200; cat. no. 11081-1-AP; ProteinTech Group, Inc.), anti-FLNa (1:200; cat. no. ab76289; Abcam), anti-P16 (1:200; cat. no. 10883-1-AP; ProteinTech Group, Inc.), and anti-Ki67 (1:1,000; cat. no. ab92742; Abcam).
The following primary antibodies were used for western blotting of relevant proteins: anti-UCP2 (1:1,000; cat. no. 11081-1-AP; ProteinTech Group, Inc.), anti-FLNa (1:1,000; cat. no. ab76289; Abcam), anti-GAPDH (1:5,000; cat. no. ab8245; Abcam), anti-ERK1/2 (1:1,000; cat. no. 4695; Cell Signaling Technology, Inc.), anti-P-ERK1/2 (1:1,000; cat. no. 4370; Cell Signaling Technology, Inc.), anti-AKT1 (1:1,000; cat. no. ab227100; Abcam), anti-P-AKT1 (1:10,000; cat. no. ab81283; Abcam), and anti-BCL-2 (1:1,000 cat. no. ab32124; Abcam).

Wang A., Liu L., Yuan M., Han S., You X., Zhang H., Lei F, & Zhang Y. (2020). Role and mechanism of FLNa and UCP2 in the development of cervical cancer. Oncology Reports, 44(6), 2656-2668.

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Analysis of BMI1 and related pathways

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Docetaxel, and PTC-209 were purchased from selleckchem (Houston, TX USA). The ChIP-grade anti-BMI1 antibody was obtained from Millipore (Bedford, MA, USA). We used following antibodies: anti-BCL2, anti-BMI1, anti-mitochondrial protein, anti-PCNA, anti-Cyclin D1 (Cell Signaling, Danvers, MA), anti-P16, anti-c-MYC (Abcam, Cambridge, MA) and anti-VEGF (SantCruz, CA). We used following plasmids pTK-TCF-Luc (Upstate Laboratories, Lake Placid, NY); pGL3-MMP2, pGL3-VEGF, pGL3-MYC (Addgene, Cambridge, MA) and pGL3-BMI1 (gifted by Goberdhan Dimri, George Washington University, Washington DC).
Transfections, luciferase reporter activity, Immunostaining, Immunoblot analysis, 3[H]-thymidine incorporation, cell growth, migration and invasion assays were performed as described previously (13 (link), 14 , 16 (link)).

Ganaie A.A., Beigh F.H., Astone M., Ferrari M.G., Maqbool R., Umbreen S., Parray A.S., Siddique H.R., Hussain T., Murugan P., Morrissey C., Koochekpour S., Deng Y., Konety B.R., Hoeppner L.H, & Saleem M. (2018). BMI1 drives metastasis of prostate cancer in Caucasian and African-American men and is a potential therapeutic target: hypothesis tested in race-specific models. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(24), 6421-6432.

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Senescence Markers Evaluation in NPCs

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Western blot was used for the expression of senescence markers P16 and P21 in NPCs in each group. According to the instructions, the total protein of each group of NPCs was extracted in RIPA lysis buffer (Millipore, Billerica, MA, USA), Equal amounts of protein were subsequently added to each well and separated by sodium dialkylsulfonate-polyacrylamide gel electrophoresis (SDS–PAGE) and then, transferred to polyvinylidene fluoride membranes (Bio-Rad Laboratories, Inc., Hercules, CA, USA). Next, the membranes were incubated with 5% bovine serum albumin for 1 h, followed by overnight incubation at 4 °C with various primary antibodies: anti-p16 (Abcam, USA), anti-P21 (Abcam, USA) and GAPDH (Ambion, Austin, TX), all primary antibodies were diluted 1:1000. Next, the membrane was washed and incubated with secondary antibody (1:8000, Bioworld technology, China) for 2 h at room temperature. Protein signals on the bands were visualized by enhanced chemiluminescence (Thermo Fisher Scientific, Waltham, USA). Protein expression levels were quantified by densitometry using ImageJ 64 software, and relative protein expression was compared to GAPDH.

Chen R., Zhang X., Zhu X., Wang C, & Xu W. (2023). Myricetin alleviated hydrogen peroxide-induced cellular senescence of nucleus pulposus cell through regulating SERPINE1. Journal of Orthopaedic Surgery and Research, 18, 143.

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Immunofluorescence Imaging of Cellular Markers

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Cell fixation and blocking were performed according to manufacturer instructions. Cells were incubated with antibodies (anti-p21, Cell Signaling Technology (Danvers, MA); anti-p16, Abcam (Cambridge, MA); anti-phospho-p38 MAPK (Thr180/Tyr182), Cell Signaling Technology; anti-γH2AX, Cell Signaling Technology; anti-GM130, BD Bioscience, Clontech, Palo Alto, CA; anti-TGN46, Sigma, St Louis, MO) and with Alexa Fluor 555-labeled secondary antibody F(ab’) fragment (Life Technologies). After washing the cells with PBS, cells were incubated with 1 μg/mL Hoechst 33342 solution for 30 min. Each well was imaged by using IN Cell Analyzer 1000, analyzed by Developer, and visualized by Spotfire DecisionSite Client 8.2 as described above.

Udono M., Fujii K., Harada G., Tsuzuki Y., Kadooka K., Zhang P., Fujii H., Amano M., Nishimura S.I., Tashiro K., Kuhara S, & Katakura Y. (2015). Impaired ATP6V0A2 expression contributes to Golgi dispersion and glycosylation changes in senescent cells. Scientific Reports, 5, 17342.

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Quantitative Proteomic and Transcriptomic Analysis

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The antibodies used in this study were as follows: anti- TERT (MBL, Nagoya, Japan), anti-p16, anti-YB-1 (Abcam, Cambridge, MA, USA), YB-1^S102p (Cell signaling Technology, Beverly, Massachusetts, USA), anti-p21-HRP (Santa Cruz, CA, USA), anti-αSMA, anti-β-actin and anti-GAPDH (Sigma, St. Louis). Proteins were visualized using an enhanced chemiluminescence system (Perkin Elmer, Waltham, MA, USA) and scanned for quantitative analysis using an Imager 600 with ImageQuant TL software (GE Healthcare, Chicago, IL, USA).
Taqman primers (Thermo Fisher Scientific) were used for qPCR analysis of human and mouse TERT, αSMA, YB-1, p16, p21, IL6, SIRT1 and 18s rRNA. One-step RT-PCR was performed as before [60 (link)] using a GeneAmp 7500 sequence detection system (Applied Biosystems, Rockford, IL). Results were expressed as 2^−ΔΔCT using the indicated control group as calibrator and 18srRNA as reference [61 (link)].

Harada M., Hu B., Lu J., Wang J., Rinke A.E., Wu Z., Liu T, & Phan S.H. (2021). The dual distinct role of telomerase in repression of senescence and myofibroblast differentiation. Aging (Albany NY), 13(13), 16957-16973.

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Western Blot Analysis of Mitochondrial Proteins

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Proteins from HUVECs were extracted with RIPA lysis buffer (Beyotime, Shanghai, China) containing 1% PMSF (Beyotime, Shanghai, China) and quantified using a BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA, USA). Equal amounts of protein (30 μg) were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), transferred to 0.22-μm PVDF membranes (Millipore, Billerica, MA, USA), and incubated overnight at 4 °C with specific antibodies: anti-p16, anti-p21, anti-FIS1, anti-SAHH (Abcam, Cambridge, UK), anti-p53, anti-Drp1, anti-OPA1, anti-MFN1, anti-MFN2, anti-DNMT1, anti-DNMT3A, anti-DNMT3B, and anti-GAPDH (Cell Signaling Technology, Danvers, MA) followed by incubation with horseradish peroxidase-conjugated secondary antibodies (Cell Signaling Technology, Danvers, MA). The dilution ratio was 1:1000 for the primary antibodies and 1:10,000 for the secondary antibodies. Protein bands were visualized with Super Signal Chemiluminescence Substrate (Thermo Scientific, Waltham, MA, USA) and GAPDH was used as a control. Images were captured using the FluorChem E system (Protein Simple, Minneapolis, MN, USA) and band densities were quantified using image J software (NIH, Bethesda, MD, USA). The densities of the bands were normalized to that of GAPDH.

You Y., Chen X., Chen Y., Pang J., Chen Q., Liu Q., Xue H., Zeng Y., Xiao J., Mi J., Tang Y, & Ling W. (2023). Epigenetic modulation of Drp1-mediated mitochondrial fission by inhibition of S-adenosylhomocysteine hydrolase promotes vascular senescence and atherosclerosis. Redox Biology, 65, 102828.

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