CRC tissue section review was performed independently by an experienced pathologist (J.-H. K.). A section of cut edge from fresh-frozen tissue was used for SA-β-Gal staining. The other section of cut edge from FFPE tissue was used for IHC analysis. IHC analysis was performed using a Benchmark XT automated processor (Ventana Medical Systems Inc, Tucson, AZ) on 4-μm-thick tissue sections of FFPE blocks. The following primary antibodies were used: p16INK4A, predilution (805–4713, Roche, Tucson, AZ); p15INK4B, 1:700 (SAB4500078, Sigma-Aldrich, Burlington, MA); p21Waf1, 1:300 (2990-1, Epitomics, Burlingame, CA); p27Kip1, 1:100 (NB110-66664, Novus Biologicals, Littleton, CO); p53, predilution (805–4713, Roche, Tucson, AZ); and p14ARF, 1:200 (74560, Cell Signaling Technology, Danvers, MA).
P15ink4b
Manufactured by Merck Group
Sourced in Germany
P15INK4B is a lab equipment product produced by Merck Group. It is a protein that functions as a cyclin-dependent kinase inhibitor, playing a role in cell cycle regulation. The core function of P15INK4B is to inhibit the activity of certain cyclin-dependent kinases, which are involved in the progression of the cell cycle.
Automatically generated - may contain errors
Lab products found in correlation
P14arf, by Cell Signaling Technology (1 mentions)
Benchmark xt, by Roche (1 mentions)
P16ink4a, by Roche (1 mentions)
Superscript 3 reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
Lightcycler 480 sybr green 1 master, by Roche (1 mentions)
Nucleospin rna kit, by Macherey-Nagel (1 mentions)
2 protocols using p15ink4b
1
Immunohistochemical Analysis of Cell Cycle Regulators
2
Quantitative gene expression analysis
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RNA was prepared using a Nucleo Spin RNA-Kit (MACHERY-NAGEL, Düren, Germany). RNA was reverse transcribed to single-stranded cDNA with SuperScriptIII Reverse Transcriptase and oligo (dT)20 50 µM (Thermo Fisher Scientific). Primer pairs for GAPDH, SMAD3 and c-MYC were obtained from Apara-bioscience GmbH (Denzlingen, Germany), p15Ink4b from Sigma-Aldrich Inc. (St. Louis, MO, USA) and p21Cip1 from Integrated DNA Technologies IDT, Inc. (Coralville, IA, USA). Primer sequences are listed in Supplementary Table S1 . Gene expression was analyzed using the LightCycler® 480 RT-qPCR and LightCycler® 480 SYBR Green I Master (Roche, Basel, Switzerland). Relative quantification of gene expression was performed as indicated. RNA sequencing and gene set enrichment analysis of generated dasatinib-resistant cells were previously described [12 (link)]. (Gene Expression Omnibus—http://ncbi.nlm.nih.gov/geo accession number GSE97352, accessed on 3 April 2017.
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